Search Results (125)

Phosphorus (P), as one of the essential bulk elements for plant growth and development, plays an important role in root growth, accumulation of secondary metabolites, and regulation of gene expression. In Salvia miltiorrhiza Bunge (S. miltiorrhiza), an important medicinal plant, the accumulation of its active components is closely related to the level of phosphorus supply, but the molecular regulatory mechanism of phosphorus treatment in the growth and secondary metabolism of S. miltiorrhiza is not clear. In this study, we investigated the effects of low phosphorus (P2), moderate phosphorus (P4), and high phosphorus (P6) treatment on the growth and development of S. miltiorrhiza. seedlings, the accumulation of bioactive compounds, and their transcriptional regulation using transcriptomic and metabolomic analyses, and identified the key regulatory genes in the biosynthesis pathways of tanshinone and salvianolic acid. The findings revealed that S. miltiorrhiza biomass exhibited a “peaked” response to phosphorus concentration, peaking at 0.625 mmol·L⁻¹. At this optimal concentration, all four batches achieved maximum root length, root weight, and leaf weight: Batch I (11.3 cm, 2.34 g, 1.62 g), Batch II (12.7 cm, 2.67 g, 1.89 g), Batch III (13.8 cm, 2.85 g, 2.04 g), and Batch IV (15.6 cm, 3.51 g, 2.44 g). Both lower and higher concentrations resulted in growth inhibition and reduced bioactive compound accumulation. Transcription factors associated with root growth and development included bHLH, MYB, and WRKY; in particular, the bZIP23 transcription factor was highly expressed under abnormal phosphorus supply conditions. In addition, the biosynthetic pathways of tanshinone and salvianolic acid were elucidated, and key genes related to the synthesis pathways (CPS, KSL, CYP, PAL, HPPR, and RAS) were identified. The expression of several TFs (such as SmCPS1, SmCYP76AH3, SmCYP76AH1, SmGGPPS1, and SmRAS1) was found to be correlated with tanshinone and salvianolic acid synthesis. The present study provides a theoretical basis for further revealing the molecular mechanism of phosphorus regulation of growth, development, and secondary metabolism of S. miltiorrhiza and provides potential targets for efficient cultivation and molecular breeding of S. miltiorrhiza. Full article

Salvia miltiorrhiza is a traditional herbal remedy for cardiovascular diseases and is in high demand in the market. Excessive chemical fertilizer application, resulting from unscientific fertilization practices, reduced the tanshinone content in S. miltiorrhiza roots. This study investigated how different fertilization types alter the endophytic microbial community composition of S. miltiorrhiza through field experiments, aiming to understand how fertilization affects its medicinal quality. The results showed that root fertilizers (F1) significantly increased root biomass and tanshinone I content, whereas foliar fertilizers (F2) increased tanshinone IIA content. High-throughput sequencing further revealed that F2 treatment significantly decreased the Shannon index of endophytic bacteria while significantly increasing the Shannon index of endophytic fungi. Co-occurrence network analysis revealed that fertilization significantly altered fungal community complexity and modularity, with F1 increasing network nodes and edges. Variance partitioning analysis indicated fungal diversity more strongly influenced medicinal compound levels under F2 and a combination of both (F3) than bacterial diversity. Septoria and Gibberella were positively correlated with tanshinone I and cryptotanshinone content under F2 treatment, respectively. Notably, the unique strains were isolated from different fertilization treatments for subsequent bacterial fertilizer development. These findings elucidate microbial responses to fertilization, guiding optimized cultivation for improved S. miltiorrhiza quality. Full article

YTH domain-containing proteins act as the primary readers of N⁶-methyladenosine (m⁶A), playing an important role in plant development and stress responses. However, little is known about the YTH proteins in medicinal plants. Genome-wide identification of the YTH gene family in the medicinal model plant, Salvia miltiorrhiza Bunge, identified a total of nineteen SmYTH genes from five chromosomes, with SmYTH8–SmYTH19 clustered on chromosome 8. Phylogenetic analysis showed that SmYTH proteins belong to the YTHDF category. No YTHDC members were identified. Conserved domain identification, amino acid sequence alignment, and phase separation prediction revealed that SmYTH1–SmYTH4 exhibited the characteristic m⁶A reader protein feature, containing conserved aromatic cages (WWW) capable of binding m⁶A residues. SmYTH5–SmYTH19 proteins contain a unique conserved F-box protein interaction domain that has not been reported previously. qRT-PCR analysis revealed tissue-specific patterns, with SmYTH1–SmYTH4 genes highly expressed in roots and leaves, whereas SmYTH8–SmYTH19 were mainly expressed in leaves. The results were consistent with RNA-seq data. The expression of various SmYTHs and the content of phenolic acid active ingredients were significantly altered under MeJA and SA treatments. The results provide useful information for further studies on the biological functions of m⁶A and YTH proteins in S. miltiorrhiza. Full article

Mining valuable genes is helpful to breed high-quality Salvia miltiorrhiza exhibiting efficient nitrogen fertilizer utilization efficiency. In the present study, transcriptome sequencing was introduced to select the candidate transcription factors (TFs) involved in tanshinones’ (TAs) and phenolic acids’ (PHAs) biosynthesis as well as low nitrogen (LN) stress. In totally, 97.71 Gb clean data was obtained from fifteen sequencing samples and 30,975 unigenes were assembled. Among of them, 27,843 unigenes were successfully annotated. Overall, 8663 differential expression genes (DEGs) were identified, among of which 5034 unigenes were up-regulated, and 3629 unigenes were down-regulated. By enrichment of DEGs together with gene co-expression network construction, 10 candidate TFs including HSFB2b, LBD12, ERF1A, ERF98, LBD25, HSF24, RAM1, HSFA4B, TCP8, and WRKY24 were finally retrieved, which are predicted to participate in modulating TA and PHA biosynthesis under LN stress. Quantitative real-time polymerase chain reaction (qRT-PCR) detection was introduced to further detect the expression profile of candidate TFs under LN stress. These findings offer a valuable resource for in-depth study of TAs ‘and PHAs’ biosynthesis under LN stress in S. miltiorrhiza. Full article

Salvia miltiorrhiza is a herbaceous plant that possesses significant medicinal value. Land salinization affects the growth of S. miltiorrhiza, resulting in a decline in its quality and yield. Knotted1-like homeobox (KNOX) genes are transcription factors that play important roles in plant growth and abiotic stress. The characteristics and functions of KNOX genes in S. miltiorrhiza remain unclear. Here, we identified ten KNOX genes in S. miltiorrhiza, all of which possess the characteristic four domains: KNOX1, KNOX2, ELK, and HD. These SmKNOXs were divided into two groups together with homologous genes. Cis-acting element analysis indicated all SmKNOXs contained elements associated with phytohormone, light, and stress response. The SmKNOXs show tissue-specific expression among roots, stems, leaves, and flowers. We assessed the response of the SmKNOXs to salt stress using quantitative RT-PCR analysis. Notably, SmKNOX4 expression significantly decreased within 24 h of salt exposure, while SmKNOX1, SmKNOX2, SmKNOX3, SmKNOX8, and SmKNOX9 showed significant increases. The expression of SmKNOX1, SmKNOX2, and SmKNOX3 was significantly positively correlated with that of their target genes, GA20ox1 and S11 MYB. These findings suggest that SmKNOXs and their target genes respond to salt stress, providing a foundation for studies of SmKNOXs and the potential genetic improvement of S. miltiorrhiza. Full article

Hair loss is one of the skin conditions that can affect people’s mental health. Plant raw material extracts are of great interest due to their safety. In this study, we utilize reverse network pharmacology to screen for key targets of the Wnt/β-catenin signaling pathway and the TGFβ/BMP signaling pathway, as well as key differential lipids, for plant raw materials selection. The aim is to identify plant raw materials that may have anti-hair loss properties and to validate these findings through cell experiments. Licorice, salvia miltiorrhiza, mulberry leaf, ephedra and curcumae radix were found that may possess anti-hair loss effects. Licorice water extract (LWE), salvia miltiorrhiza water extract (SMWE), mulberry leaf water extract (MLWE), ephedra water extract (EWE) and curcumae radix water extract (CRWE) did not exhibit cytotoxicity on human dermal papilla cells (HDPCs). Through ALP staining, it was found that the expression of ALP in HDPCs treated with LWE, SMWE, MLWE, EWE and CRWE was enhanced. In addition, LWE, SMWE, MLWE, EWE and CRWE have reduced the expression of hair growth inhibitory factor TGF-β1 and inflammatory factor IL-6. Additionally, various water extracts can enhance the secretion of VEGF, with high concentrations of SMWE, EWE and CRWE exhibiting better efficacy. Furthermore, β-catenin, a key factor of the Wnt/β-catenin signaling pathway, was enhanced by LWE, SMWE, MLWE, EWE and CRWE treatment in cultured HDPCs. In conclusion, all five plant raw materials showed some anti-hair loss potential, providing theoretical support for their application in anti-hair loss products. Full article

Tanshinones, biologically active diterpene compounds derived from Salvia miltiorrhiza, interact with specific proteins and DNA sequences, influencing signaling pathways in animals and humans. This study highlights tanshinone–protein interactions observed at concentrations achievable in vivo, ensuring greater physiological relevance compared to in vitro studies that often employ supraphysiological ligand levels. Experimental data suggest that while tanshinones interact with multiple proteomic targets, only a few enzymes are significantly affected at biologically relevant concentrations. This apparent paradox may be resolved by tanshinones’ ability to bind DNA and influence enzymes involved in gene expression or mRNA stability, such as RNA polymerase II and human antigen R protein. These interactions trigger secondary, widespread changes in gene expression, leading to complex proteomic alterations. Although the current understanding of tanshinone–protein interactions remains incomplete, this study provides a foundation for deciphering the molecular mechanisms underlying the therapeutic effects of S. miltiorrhiza diterpenes. Additionally, numerous tanshinone derivatives have been developed to enhance pharmacokinetic properties and biological activity. However, their safety profiles remain poorly characterized, limiting comprehensive insights into their medicinal potential. Further investigation is essential to fully elucidate the therapeutic and toxicological properties of both native and modified tanshinones. Full article

Salvia miltiorrhiza Bunge (S. miltiorrhiza) is one of the most commonly used bulk herbs in China; however, root rot can seriously affect its quality and yields. To minimize the use of chemical pesticides for managing this plant fungal disease, biological control utilizing microbial bio-pesticides offers a promising alternative. This study aimed to enhance the biocontrol resources available for combating S. miltiorrhiza root rot by isolating actinomycetes with antifungal activity from the rhizosphere soil of S. miltiorrhiza and identifying biocontrol actinomycetes with a preventive effect on root rot. A total of 35 actinomycetes were successfully screened from the rhizosphere soil of S. miltiorrhizae. The strain B-35 with the strongest antifungal activity was screened out through antagonizing the pathogen Fusarium solani of S. miltiorrhiza, strain morphology and 16S rRNA analysis. The antagonistic actinomycetes fermentation filtrate and crude extract could significantly destroy the mycelium and spores of Fusarium solani; the biocontrol effect of mature S. miltiorrhiza rhizome reached 83.3%, and the number of leaves, plant height and biomass in the B-35 treatment group were significantly increased compared with the control group. B-35 has a certain application potential in the biological control of root rot and the promotion of S. miltiorrhizae. The antifungal activity of actinomycetes sourced from the rhizosphere soil of S. miltiorrhiza has been demonstrated for the first time, potentially enhancing future crop quality and production. Full article

Salvia miltiorrhiza, the valuable traditional Chinese medicinal plant, has been used in clinics for thousands of years. The water-soluble salvianolic acid compounds are bioactive substances used in treating many diseases. Gibberellins (GAs) are growth-promoting phytohormones that regulate plant growth and development. Previous studies have demonstrated that GAs can promote salvianolic acid accumulation in S. miltiorrhiza; however, the underlying mechanism requires further investigation. Here, we identified a GA-induced R2R3MYB transcription factor (TF), SmMYB71, from a transcriptome library of GA-treated S. miltiorrhiza. SmMYB71 was highly expressed in the root of S. miltiorrhiza and localized to the nucleus. SmMYB71-knockout hairy roots showed higher salvianolic acid accumulation compared to wild lines. Overexpressing SmMYB71 in S. miltiorrhiza hairy roots significantly decreased the content of salvianolic acid by downregulating key salvianolic acid biosynthesis enzymes such as SmRAS and SmCYP98A14. The GCC box in the promoter of SmMYB71 can bind with SmERF115, suggesting that SmMYB71 is regulated by SmERF115 in salvianolic acid biosynthesis. These findings demonstrate a novel regulatory role of SmMYB71 in GA-mediated phenolic acid biosynthesis. With the development of CRISPR/Cas9-based genome editing technology, the SmMYB71 regulation mechanism of salvianolic acid biosynthesis provides a potential target gene for metabolic engineering to increase the quality of S. miltiorrhiza. Full article

Danshen (Salvia miltiorrhiza Bunge) is a perennial herbaceous plant of the Salvia genus in the family Lamiaceae. Its dry root is one of the important traditional Chinese herbal medicines with a long officinal history. The yield and quality of S. miltiorrhiza are influenced by various factors, among which drought is one of the most significant types of abiotic stress. Based on the transcriptome database of S. miltiorrhiza, our research group discovered a carotenoid cleavage dioxygenase gene, SmCCD4, belonging to the carotenoid cleavage oxygenase (CCO) gene family which is highly responsive to drought stress on the basis of our preceding work. Here, we identified 26 CCO genes according to the whole-genome database of S. miltiorrhiza. The expression pattern of SmCCD4 showed that this gene is strongly overexpressed in the aboveground tissue of S. miltiorrhiza. And by constructing SmCCD4 overexpression strains, it was shown that the overexpression of SmCCD4 not only promotes the synthesis of abscisic acid and increases plant antioxidant activity but also regulates the synthesis of the secondary metabolites tanshinone and phenolic acids in S. miltiorrhiza. In summary, this study is the first in-depth and systematic identification and investigation of the CCO gene family in S. miltiorrhiza. The results provide useful information for further systematic research on the function of CCO genes and provide a theoretical basis for improving the yield and quality of S. miltiorrhiza. Full article

In the process of catalyzing carotenoids into various apocarotenoids and other derivatives, carotenoid cleavage dioxygenases (CCDs) play key roles. However, little information on CCDs has been reported in regard to Salvia miltiorrhiza. In this study, a total of 21 CCD genes were identified in the whole genome of S. miltiorrhiza, mainly distributed between five chromosomes. Phylogenetic relationship analysis revealed that 21 SmCCD genes were classified into four subfamilies, including SmCCD4, 7, 8, and NCED; the members of the same subfamily show similar gene structures and tertiary structures. The interspecific collinearity with other plant species, such as Arabidopsis thaliana and Oryza sativa was analyzed. Cis-elements analysis demonstrated that the majority were stress response-, light response-, growth-, and development-related. The expression pattern of the SmCCD genes was expressed in the analyzed tissues. Furthermore, the majority of the SmCCD4 subfamily members varied in their expression levels under the treatment of MeJA, YE, and ABA, indicating the potential function of SmCCD4 in the metabolism process of S. miltiorrhiza. In general, this study provides a systematic analysis of SmCCD genes and lays the foundation for uncovering the regulation and function of SmCCD genes in S. miltiorrhiza. Full article

Phenolic acids have health-promoting properties, however, but their low concentrations in Salvia miltiorrhiza limit broader medicinal applications. MYB and bHLH transcription factors activate multiple target genes involved in phenylpropanoid metabolism, thereby enhancing the production of various secondary metabolites. We introduced the MYB transcription factor Antirrhinum Rosea1 (AmROS1) or Delila (AmDEL) into S. miltiorrhiza and observed that antioxidant activity in transgenic plants increased by 1.40 to 1.80-fold. The total content was significantly higher in transformants compared to the controls. Furthermore, heterologous expression of AmROS1 or AmDEL triggered moderate accumulations of rosmarinic acid and salvianolic acid at various growth stages. Levels of total phenolics, total flavonoids, and anthocyanins were significantly elevated. These biological and phytochemical alterations were correlated with the upregulated expression of genes involved in phenolic acid biosynthesis. Our findings demonstrate that AmROS1 and AmDEL function as a transcriptional activator in phenolic acids biosynthesis. This study offers further insights into the heterologous or homologous regulation of phenolics production, potentially enabling its engineering in S. miltiorrhiza. Full article

The root of Salvia miltiorrhiza Bunge (SMB) has been widely used to treat cardiovascular diseases. However, the contents of secondary metabolites in the roots from different production areas are significantly different, and the impact of soil factors on this accumulation remains unclear. Therefore, this study aimed to elucidate the regularity of variation between the active components and soil factors through targeted metabolomics and chemical dosimetry. Soils were collected from five different cities (A, B, C, D, and E) and transplanted into the study area. The results showed that there were significant differences in the soil fertility characteristics and heavy metal pollution levels in different soils. Ten water- and twelve lipid-soluble metabolites were identified in SMBs grown in all soil types. SMBs from D cities exhibited the highest total tanshinone content (p < 0.05). The salvianolic acid B content in SMBs from E cities was the highest (p < 0.05). Interestingly, correlation analysis revealed a significant negative correlation between the accumulation of lipid-soluble and water-soluble metabolites. Double-matrix correlation analysis demonstrated that available potassium (AK) was significantly negatively correlated with salvianolic acid B (r = −0.80, p = 0.0004) and positively correlated with tanshinone IIA (r = 0.66, p = 0.008). Conversely, cadmium (Cd) and cuprum (Cu) were significantly positively and negatively correlated with salvianolic acid B (r = 0.96, p < 0.0001 and r = 0.72, p = 0.0024) and tanshinone IIA (r = 0.40, p = 0.14 and r = 0.73, p = 0.0018), respectively. Mantel’s test indicated that AK (r > 0.52, p < 0.001), Cu (r > 0.60, p < 0.005), and Cd (r > 0.31, p < 0.05) were the primary drivers of the differences in the active components of SMBs. These findings provide a theoretical framework for modulating targeted metabolites of SMB through soil factors, with significant implications for the cultivation and quality control of medicinal plants. Full article

Diabetic nephropathy, a leading cause of end-stage renal disease, accounts for significant morbidity and mortality. It is characterized by microinflammation in the glomeruli and myofibroblast activation in the tubulointerstitium. Salvia miltiorrhiza Bunge, a traditional Chinese medicine, is shown to possess anti-inflammatory and anti-fibrotic properties, implying its renal-protective potential. This study investigates which type of component can reduce the damage caused by diabetic nephropathy in a single setting. The ethyl acetate (EtOAc) layer was demonstrated to provoke peroxisome proliferator-activated receptor (PPAR)-α and PPAR-γ activities in renal mesangial cells by dual luciferase reporter assay. In a high glucose (HG)-cultured mesangial cell model, the EtOAc layer substantially inhibited HG-induced elevations of interleukin-1β, transforming growth factor-β1 (TGF-β1), and fibronectin, whereas down-regulated PPAR-γ was restored. In addition, among the extracts of S. miltiorrhiza, the EtOAc layer effectively mitigated TGF-β1-stimulated myofibroblast activation. The EtOAc layer also showed a potent ability to attenuate renal hypertrophy, proteinuria, and fibrotic severity by repressing diabetes-induced proinflammatory factor, extracellular matrix accumulation, and PPAR-γ reduction in the STZ-induced diabetes mouse model. Our findings, both in vitro and in vivo, indicate the potential of the EtOAc layer from S. miltiorrhiza for future drug development targeting diabetic nephropathy. Full article

Hyaluronidase possesses the capacity to degrade high-molecular-weight hyaluronic acid into smaller fragments, subsequently initiating a cascade of inflammatory responses and activating dendritic cells. In cases of bacterial infections, substantial quantities of HAase are generated, potentially leading to severe conditions such as cellulitis. Inhibiting hyaluronidase activity may offer anti-inflammatory benefits. Salvia miltiorrhiza Bunge, a traditional Chinese medicine, has anti-inflammatory properties. However, its effects on skin inflammation are not well understood. This study screened and evaluated the active components of S. miltiorrhiza that inhibit skin inflammation, using ligand fishing, enzyme activity assays, drug combination analysis, and molecular docking. By combining magnetic nanomaterials with hyaluronidase functional groups, we immobilized hyaluronidase on magnetic nanomaterials for the first time in the literature. We then utilized an immobilized enzyme to specifically adsorb the ligand; two ligands were identified as salvianolic acid B and rosmarinic acid by HPLC analysis after desorption of the dangling ligands, to complete the rapid screening of potential anti-inflammatory active ingredients in S. miltiorrhiza roots. The median-effect equation and combination index results indicated that their synergistic inhibition of hyaluronidase at a fixed 3:2 ratio was enhanced with increasing concentrations. Kinetic studies revealed that they acted as mixed-type inhibitors of hyaluronidase. Salvianolic acid B had K_i and K_is values of 0.22 and 0.96 μM, respectively, while rosmarinic acid had values of 0.54 and 4.60 μM. Molecular docking revealed that salvianolic acid B had a higher affinity for hyaluronidase than rosmarinic acid. In addition, we observed that a 3:2 combination of SAB and RA significantly decreased the secretion of TNF-α, IL-1, and IL-6 inflammatory cytokines in UVB-irradiated HaCaT cells. These findings identify salvianolic acid B and rosmarinic acid as key components with the potential to inhibit skin inflammation, as found in S. miltiorrhiza. This research is significant for developing skin inflammation treatments. It demonstrates the effectiveness and broad applicability of the magnetic nanoparticle-based ligand fishing approach for screening enzyme inhibitors derived from herbal extracts. Full article