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The Panax L. genus is well-known for many positive physiological effects on humans, with major species including P. ginseng , P. quinquefolius , P. notoginseng , P. japonicus , and P. japonicus var. major , the first three of which are globally popular. The combination of UPLC-QTOF-MS and chemometrics were developed to profile "identification markers" enabling their differentiation. The establishment of reliable biomarkers that embody the intrinsic metabolites differentiating species within the same genus is a key in the modernization of traditional Chinese medicine. In this work, the metabolomic differences among these five species were shown, which is critical to ensure their appropriate use. Consequently, 49 compounds were characterized, including 38 identified robust biomarkers, which were mainly composed of saponins and contained small amounts of amino acids and fatty acids. VIP (projection variable importance) was used to identify these five kinds of ginseng. In conclusion, by illustrating the similarities and differences between the five species of ginseng with the use of an integrated strategy of combining UPLC-QTOF-MS and multivariate analysis, we provided a more efficient and more intelligent manner for explaining how the species differ and how their secondary metabolites affect this difference. The most important biomarkers that distinguished the five species included Notoginsenoside-R 1 , Majonoside R 1 , Vinaginsenoside R 14 , Ginsenoside-Rf, and Ginsenoside-Rd.

The leaves of Panax species (e.g., Panax ginseng -PGL, P. quinquefolius -PQL, and P. notoginseng -PNL) can serve as a source for healthcare products. Comprehensive characterization and unveiling of the metabolomic difference among PGL, PQL, and PNL are critical to ensure their correct use. For this purpose, enhanced profiling and chemometrics were integrated to probe into the ginsenoside markers for PGL/PQL/PNL by ultra-high performance liquid chromatography/ion mobility-quadrupole time-of-flight mass spectrometry (UHPLC/IM-QTOF-MS). A hybrid scan approach (HDMS E -HDDDA) was established achieving the dimension-enhanced metabolic profiling, with 342 saponins identified or tentatively characterized from PGL/PQL/PNL. Multivariate statistical analysis (33 batches of leaf samples) could unveil 42 marker saponins, and the characteristic ginsenosides diagnostic for differentiating among PGL/PQL/PNL were primarily established. Compared with the single DDA or DIA, the HDMS E -HDDDA hybrid scan approach could balance between the metabolome coverage and spectral reliability, leading to high-definition MS spectra and the additional collision-cross section (CCS) useful to differentiate isomers.

Panax ginseng (PG) and Panax notoginseng (PN) are highly valuable Chinese medicines (CM). Although both CMs have similar active constituents, their clinical applications are clearly different. Over the past decade, RNA sequencing (RNA-seq) analysis has been employed to investigate the molecular mechanisms of extracts or monomers. However, owing to the limited number of samples in standard RNA-seq, few studies have systematically compared the effects of PG and PN spanning multiple conditions at the transcriptomic level. Here, we developed an approach that simultaneously profiles transcriptome changes for multiplexed samples using RNA-seq (TCM-seq), a high-throughput, low-cost workflow to molecularly evaluate CM perturbations. A species-mixing experiment was conducted to illustrate the accuracy of sample multiplexing in TCM-seq. Transcriptomes from repeated samples were used to verify the robustness of TCM-seq. We then focused on the primary active components, Panax notoginseng saponins (PNS) and Panax ginseng saponins (PGS) extracted from PN and PG, respectively. We also characterized the transcriptome changes of 10 cell lines, treated with four different doses of PNS and PGS, using TCM-seq to compare the differences in their perturbing effects on genes, functional pathways, gene modules, and molecular networks. The results of transcriptional data analysis showed that the transcriptional patterns of various cell lines were significantly distinct. PGS exhibited a stronger regulatory effect on genes involved in cardiovascular disease, whereas PNS resulted in a greater coagulation effect on vascular endothelial cells. This study proposes a paradigm to comprehensively explore the differences in mechanisms of action between CMs based on transcriptome readouts. [Display omitted] • A low-cost workflow of high-throughput transcriptional profiling was developed. • The atlas of signatures perturbed by PGS and PNS was established using TCM-seq. • The regulation of PGS and PNS was holistically compared with network analysis. • A paradigm of molecularly evaluating CMs with transcriptome readouts was proposed. [ABSTRACT FROM AUTHOR]

Porcine circovirus type 2 (PCV2) exists widely in swine populations worldwide, and healthy PCV2 virus carriers have enhanced the severity of the infection, which is becoming more difficult to control. This study investigated the regulatory effect of Panax notoginseng saponins (PNS) on the oxidative stress and histone acetylation modification induced by PCV2 in vitro and in mice. In vitro, PNS significantly increased the scavenging capacities of superoxide anion radicals (O2 •-) and hydroxyl radicals (• OH) and reduced the content of hydrogen peroxide (H2O2) induced by PCV2 in porcine alveolar macrophages (3D4/2). In addition, PNS decreased the protein expression level of histone H4 acetylation (Ac-H4) by increasing the activity of histone deacetylase (HDAC) in PCV2-infected 3D4/2 cells. In vivo, PNS enhanced the scavenging capacities of • OH and O2 •− and reduced the content of H2O2 in the spleens of PCV2-infected mice. PNS also reduced the protein expression level of histone H3 acetylation (Ac-H3) by reducing the activity of histone acetylase (HAT) and increasing the activity of HDAC in the spleens of PCV2-infected mice. PCV2 infection activated oxidative stress and histone acetylation in vitro and in mice, but PNS ameliorated this oxidative stress. The research can provide experimental basis for exploring the antioxidant effect and the regulation of histone acetylation of PNS on PCV2-infected 3D4/2 cells and mice in vitro and in vivo, and provide new ideas for the treatment of PCV2 infection. [ABSTRACT FROM AUTHOR]

Astragali Radix (AR) is one of the famous traditional Chinese medicines (TCMs) for boosting immunity, whereas the quality markers (Q-markers) of AR have not been clearly researched. The immunomodulatory activities of the bioactive extractions and components were evaluated by NO inhibition rate; phagocytic index; IL-10, TNF-α, IL-1β, and IL-6 cytokines in RAW264.7 cells; and the relative proliferation rate of spleen cells. The total saponins (TS) and the grade 2 (Xiaoxuan, XX) of AR showed the strongest immunomodulatory activities. At the concentration of 40 μg/mL, the TS increased spleen cells proliferation by 48.0% and upregulated the level of IL-1β and IL-6. Cytokines in the XX-treated group were at least 1.6 times higher than the control group. A total of 190 common peaks were detected in AR by ultrahigh-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UHPLC-QTOF-MS). The multivariate statistical analyses revealed that 41 compounds were positively correlated with immune responses, and bioactive compounds were verified by using RAW264.7 cell assay. Subsequently, the contents of six compounds in different commercial grades were determined, and the results showed the same trend in contents and activities. Finally, calycosin-7-O-β-D-glucoside, astragaloside IV, astragaloside II, astragaloside I, isomucronulatol-7-O-glucoside, and 9,10-dimethoxypterocarpan-3-O-glucoside were screened out as immunomodulatory Q-markers of AR., (© 2024 John Wiley & Sons Ltd.)

Background: Panax notoginseng saponins (PNS) are bioactive substances extracted from P. notoginseng that are widely used to treat cardiovascular and cerebrovascular diseases and interstitial diseases. PNS have the functions of scavenging free radicals, anti-inflammation, improving blood supply for tissue and so on. Objectives: The aim of this study was to investigate the effects of PNS on the oxidative stress of immune cells induced by porcine circovirus 2 (PCV2) infection in vitro and in vivo. Methods: Using an oxidative stress model of PCV2 infection in a porcine lung cell line (3D4/2 cells) and mice, the levels of nitric oxide (NO), reactive oxygen species (ROS), total glutathione (T-GSH), reduced glutathione (GSH), and oxidized glutathione (GSSG) and the activities of xanthine oxidase (XOD), myeloperoxidase (MPO) and inducible nitric oxide synthetase (iNOS) were determined to evaluate the regulatory effects of PNS on oxidative stress. Results: PNS treatment significantly reduced the levels of NO and ROS, the content of GSSG and the activities of XOD, MPO, and iNOS (p < 0.05), while significantly increasing GSH and the ratio of GSH/GSSG in infected 3D4/2 cells (p < 0.05). Similarly, in the in vivo study, PNS treatment significantly decreased the level of ROS in spleen lymphocytes of infected mice (p < 0.05), increased the levels of GSH and T-GSH (p < 0.05), significantly decreased the GSSG level (p < 0.05), and decreased the activities of XOD, MPO, and iNOS. Conclusions: PNS could regulate the oxidative stress of immune cells induced by PCV2 infection in vitro and in vivo. [ABSTRACT FROM AUTHOR]

Abstract Panax notoginseng saponins (PNS) are the major components of Panax notoginseng , with multiple pharmacological activities but poor oral bioavailability. PNS could be metabolized by gut microbiota in vitro , while the exact role of gut microbiota of PNS metabolism in vivo remains poorly understood. In this study, pseudo germ-free rat models were constructed by using broad-spectrum antibiotics to validate the gut microbiota-mediated transformation of PNS in vivo. Moreover, a high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) was developed for quantitative analysis of four metabolites of PNS, including ginsenoside F1 (GF1), ginsenoside Rh2 (GRh2), ginsenoside compound K (GCK) and protopanaxatriol (PPT). The results showed that the four metabolites could be detected in the control rat plasma, while they could not be determined in pseudo germ-free rat plasma. The results implied that PNS could not be biotransformed effectively when gut microbiota was disrupted. In conclusion, gut microbiota plays an important role in biotransformation of PNS into metabolites in vivo. [ABSTRACT FROM AUTHOR]

Cisplatin is a major antineoplastic drug that is used to treat solid tumors, but its use is restricted by its nephrotoxicity. Such cisplatin‐induced nephrotoxicity (CIN) is believed to occur primarily through mitochondrial damage and reactive oxygen species (ROS) generation. Our previous studies have indicated that Panax notoginseng saponins (PNSs) mitigate CIN by enhancing hypoxia‐inducible factor 1α (HIF‐1α)‐induced mitochondrial autophagy. In this study, the role of the HIF‐1α/mitochondria/ROS pathway in PNSs protection against CIN was investigated using a rat model. A CIN model was generated by giving rats intraperitoneal injections with cisplatin (a single dose) and then treating them with or without 2‐methoxyestradiol (HIF‐1α inhibitor) and PNSs. We then measured ROS levels, superoxide dismutase, glutathione, catalase malondialdehyde and nitric oxide (to evaluate oxidative stress) and ATP, mitochondrial membrane potential and mitochondrial permeability transition pore opening (to evaluate mitochondrial function) in kidneys at different time points. We observed that PNSs remarkably reduced the levels of ROS, malondialdehyde and nitric oxide, as well as the opening of mitochondrial permeability transition pore, which is increased by cisplatin and further increased by HIF‐1α inhibition. In addition, PNSs increased the levels of superoxide dismutase, catalase and glutathione, as well as ATP and mitochondrial membrane potential in renal tissues; these are all reduced by cisplatin and further reduced by HIF‐1α inhibition. In conclusion, we demonstrate here that PNSs protects against mitochondrial damage induced by cisplatin through HIF‐1α/mitochondria/ROS. [ABSTRACT FROM AUTHOR]

Panax notoginseng saponins (PNS) are a commonly used traditional medicine to treat diabetes in China. Recent studies have confirmed their anti‐diabetic effects, but the underlying mechanisms have remained unclear. The present study was designed to explore whether PNS decrease hyperglycemia by improving insulin sensitivity in skeletal muscle and to elucidate the molecular mechanisms. The anti‐diabetic effects of PNS were analyzed in a skeletal myoblast cell line, C2C12, and in high fat diet‐induced diabetic KKAy mice. C2C12 cells were treated with PNS (50, 100, and 200 μg·L−1) and examined for glucose uptake, cell viability and expression of components of the phosphoinositide 3‐kinase (PI3K)–protein kinase B (AKT) signaling pathway. KKAy mice were intraperitoneally injected with PNS (200 mg·kg−1) for 6 weeks. Body weight, blood glucose, serum insulin, serum lipid, glucose and insulin tolerance were measured to evaluate the anti‐diabetic effects of PNS. Pathological changes, apoptosis and the PI3K–AKT signaling pathway were analyzed in KKAy skeletal muscle. PNS significantly increased insulin‐induced glucose uptake, but did not affect the cell viability of C2C12 cells. In addition, PNS reduced blood glucose and serum insulin levels and improved glucose tolerance and insulin tolerance of KKAy mice. Pathological changes and apoptosis of skeletal muscle were relieved by PNS treatment. Moreover, PNS treatment enhanced expression of mRNA encoding IRS1 and GLUT4, as well as the protein expression of phosphorylated (p) ‐insulin receptor substrate 1 (IRS1), p‐PI3K, p‐AKT and glucose transporter type 4 (GLUT4) in C2C12 and KKAy mouse muscle. Collectively, these data indicate that PNS reduces hyperglycemia and insulin resistance through up‐regulating GLUT4 expression and the IRS1–PI3K–AKT signaling pathway. Furthermore, PNS alleviated diabetes skeletal muscle pathological damage. Thus, our data suggest that PNS may be promising anti‐diabetic compounds. [ABSTRACT FROM AUTHOR]

Current evidence suggests a beneficial role of herbal products in free radical-induced diseases. Panax notoginseng (Burk.) F. H. Chen has long occupied a leading position in traditional Chinese medicine because of the ergogenic, nootropic, and antistress activities, although these properties are also acknowledged in the Western world. The goal of this paper is to review the pharmacological and toxicological properties of P. notoginseng and discuss its potential therapeutic effect. A literature search was carried out on Pubmed, Scopus, and the Cochrane Central Register of Controlled Trials databases. The following search terms were used: “notoginseng”, “gut microbiota”, “immune system”, “inflammation”, “cardiovascular system”, “central nervous system”, “metabolism”, “cancer”, and “toxicology”. Only peer-reviewed articles written in English, with the full text available, have been included. Preclinical evidence has unraveled the P. notoginseng pharmacological effects in immune-inflammatory, cardiovascular, central nervous system, metabolic, and neoplastic diseases by acting on several molecular targets. However, few clinical studies have confirmed the therapeutic properties of P. notoginseng, mainly as an adjuvant in the conventional treatment of cardiovascular disorders. Further clinical studies, which both confirm the efficacy of P. notoginseng in free radicalrelated diseases and delve into its toxicological aspects, are mandatory to broaden its therapeutic potential. [ABSTRACT FROM AUTHOR]

Panax notoginseng is famous for its important therapeutic effects. Saponins are bioactive compounds found in different parts and developmental stages of P. notoginseng plants. Thus, it is urgently to study saponins distribution in different parts and growth ages of P. notoginseng plants. In this study, potential biomarkers were found, and their chemical characteristic differences were revealed through metabolomic analysis. High-performance liquid chromatography data indicated the higher content of saponins ( i.e. , Rg1, Re, Rd, and Rb1) in the underground parts than that in the aerial parts. 20( S )-Protopanaxadiol saponins were mainly distributed in the aerial parts. Additionally, the total saponin content in the 3-year-old P. notoginseng plant (188.0 mg/g) was 1.4-fold higher than that in 2-year-old plant (130.5 mg/g). The transcriptomic analysis indicated the tissue-specific transcription expression of genes, namely, PnFPS , PnSS , PnSE1 , PnSE2 , and PnDS , which encoded critical synthases in saponin biosyntheses. These genes showed similar expression patterns among the parts of P. notoginseng plants. The expression levels of these genes in the flowers and leaves were 5.2fold higher than that in the roots and fibrils. These results suggested that saponins might be actively synthesized in the aerial parts and transformed to the underground parts. This study provides insights into the chemical and genetic characteristics of P. notoginseng to facilitate the synthesis of its secondary metabolites and a scientific basis for appropriate collection and rational use of this plant. [ABSTRACT FROM AUTHOR]

Objective To evaluate the antitumor activity both in vitro and in vivo of saponin–phospholipid complex of Panax notoginseng . Methods The in vitro cytotoxic effect of saponins extract and saponin–phospholipid complex against human lung cancer NCI-H460 and breast cancer cell lines BT474 was examined using MTS assay. For in vivo evaluation of antitumor potential, saponin and saponin–phospholipid complex were administered orally in rats induced mammary carcinogenesis by 7,12-dimethylbenz(a)anthracene, for 30 days. Results Our data showed that saponin–phospholipid complex had stronger anticancer effect compared to saponin extract. The IC50 values of saponin–phospholipid complex and saponin extract for NCI-H460 cell lines were 28.47 μg/mL and 47.97 μg/mL, respectively and these values for BT474 cells were 53.18 μg/mL and 86.24 μg/mL, respectively. In vivo experiments, administration of saponin, saponin–phospholipid complex and paclitaxel (positive control) effectively suppressed 7,12-dimethylbenz(a) anthracene-induced breast cancer evidenced by a decrease in tumor volume, the reduction of lipid peroxidation level and increase in the body weight, and elevated the enzymatic antioxidant activities of superoxide dismutase, catalase, glutathione peroxidase in rat breast tissue. Conclusions Our study suggests that saponin extract from Panax notoginseng and saponin–phospholipid complex have potential to prevent cancer, especially breast cancer. [ABSTRACT FROM AUTHOR]

Background We aimed to study the effect of Panax notoginseng saponins (PNS) on the proliferation, differentiation, self-renewal, and expressions of basic fibroblast growth factor (bFGF) and brain-derived neurotrophic factor (BDNF) in rat embryonic neural stem cells (NSCs). Methods Cortical stem cells were isolated from rat embryos on Embryonic Day 17 (E17) and identified by nestin expression. Subsequently, primary culture, subculturing, and single cell cloning were performed on the cells. After the first cell passage (P1), the cells were resuspended and divided into a control group and a treatment group. Control cells were cultured in serum-free basal culture medium with B27 and dulbecco's modified eagle medium (DMEM)/F12. The same medium supplemented with PNS (100 μg/mL) was used to culture cells in the treatment group. Both groups were incubated at 37°C in a 5% CO 2 incubator. Immunocytochemistry was performed 4 days after incubation. Results Primary, P1, and P2 cells in the treatment group formed neurospheres, as did single cell clones of the P1 cells in this group. After being cultured for 4 days, the number of nestin-, proliferating cell nuclear antigen (PCNA)-, Tuj-1-, neurofilament (NF)-, vimentin-, glial fibrillary acidic protein (GFAP)-, bFGF-, and BDNF-positive cells significantly increased in the treatment group in comparison to the control group. All positively stained cells could form clear clusters. Conclusion PNS can promote rat embryonic cortical NSC survival, self-renewal, proliferation, and differentiation through neurotrophic factors by autocrine or paracrine signaling. [ABSTRACT FROM AUTHOR]

Skeletal muscle is an important organ for controlling the development of type 2 diabetes. We discoveredPanax notoginsengroots as a candidate to improve hyperglycemia throughin vitromuscle cells screening test. Saponins are considered as the active ingredients of ginseng. However, in the body, saponins are converted to dammarane-type triterpenes, which may account for the anti-hyperglycemic activity. We developed a method for producing a dammarane-type triterpene extract (DTE) fromPanax notoginsengroots and investigated the extract’s potential anti-hyperglycemic activity. We found that DTE had stronger suppressive activity on blood glucose levels than the saponin extract (SE) did in KK-Aymice. Additionally, DTE improved oral glucose tolerance, insulin sensitivity, glucose uptake, and Akt phosphorylation in skeletal muscle. These results suggest that DTE is a promising agent for controlling hyperglycemia by enhancing glucose uptake in skeletal muscle. DTM improved hyperglycemia. [ABSTRACT FROM PUBLISHER]

Abstract: Eight darmarane-type saponins, named as sanchirhinoside D (1), 3β,6α-20(S)-6,20-bis(β-d-glucopyranosyloxy)-3-hydroxy dammar-24-en-12-one (2), 20(R)-ginsenoside Rg3 (3), ginsenosides F2 (4) and Rd (5), gypenoside XVII (6), ginsenosides Rb1 (7) and ginsenoside V (8) were isolated from the 70% ethanol extract of Panax notoginseng roots by silica gel, octadecyl silane (ODS) column chromatography and HPLC, and identified by chemical and physical methods, especially by spectral analysis. Among them, 1 was a new compound and 2 was a new natural product. [Copyright &y& Elsevier]

Background: Panax notoginseng (Burk) F. H. Chen is one of the most famous Chinese traditional medicinal plants. The taproot is the main organ producing triterpenoid saponins, and its development is directly linked to the quality and yield of the harvested P. notoginseng. However, the mechanisms underlying the dynamic metabolic changes occurring during taproot development of P. notoginseng are unknown. Results: We carried out metabolomic and transcriptomic analyses to investigate metabolites and gene expression during the development of P. notoginseng taproots. The differentially accumulated metabolites included amino acids and derivatives, nucleotides and derivatives, and lipids in 1-year-old taproots, flavonoids and terpenoids in 2- and 3-year-old taproots, and phenolic acids in 3-year-old taproots. The differentially expressed genes (DEGs) are related to phenylpropanoid biosynthesis, metabolic pathway and biosynthesis of secondary metabolites at all three developmental stages. Integrative analysis revealed that the phenylpropanoid biosynthesis pathway was involved in not only the development of but also metabolic changes in P. notoginseng taproots. Moreover, significant accumulation of triterpenoid saponins in 2- and 3-year-old taproots was highly correlated with the up-regulated expression of cytochrome P450s and uridine diphosphate-dependent glycosyltransferases genes. Additionally, a gene encoding RNase-like major storage protein was identified to play a dual role in the development of P. notoginseng taproots and their triterpenoid saponins synthesis. Conclusions: These results elucidate the molecular mechanism underlying the accumulation of and change relationship between primary and secondary metabolites in P. notoginseng taproots, and provide a basis for the quality control and genetic improvement of P. notoginseng. [ABSTRACT FROM AUTHOR]

Abstract: Total saponins extracted from the caudexes and leaves of Panax notoginseng (SCLPN) have been used in the clinic for improving mental function, treating insomnia, and alleviating anxiety. The present study evaluated the potential antidepressant activity of SCLPN in rodent models of depression and the mechanism of action of SCLPN. Mice were received SCLPN at doses of 10–1000mg/kg daily for 1, 7, and 14 days and then were subjected to the forced swim test and locomotor activity test. The results showed that SCLPN decreased immobility time in the forced swim test, with little effect on locomotion. In the chronic mild stress model, chronic SCLPN treatment (70mg/kg) reversed the rats’ depression-like behavior. Furthermore, SCLPN exerted its antidepressant-like effect by increasing the levels of 5-hydroxytryptamine, dopamine, and noradrenaline. Additionally, SCLPN treatment reduced intracellular Ca2+ in cultured neurons. The present study suggests that SCLPN may function as an antidepressant, and the antidepressant-like effects of SCLPN may be mediated by modulation of brain monoamine neurotransmitters and intracellular Ca2+ concentration. [Copyright &y& Elsevier]

Abstract: Objective: To investigate the effects of Panax notoginseng saponins (PNS) on hydrogen peroxide (H2O2)-induced apoptosis in cultured rabbit bone marrow stromal cells (BMSCs). Methods: The effects of different concentrations of PNS on proliferation and early osteoblast differentiation of BMSCs were determined by the MTT assay and an alkaline phosphatase (ALP) assay. An optimal effective concentration of PNS was determined and used in subsequent experiments. The cultured BMSCs were divided into three groups: untreated control, H2O2 treated, and PNS pretreatment of H2O2 treated. The oxidative stress level was assessed by superoxide dismutase (SOD) and malondialdehyde (MDA) assays. Flow cytometry was used to determine BMSC apoptosis by staining with annexinV-FITC/propidium iodide (PI). The activity of caspase-3 enzyme was measured by spectrofluorometry. Results: PNS (0.1g/L) significantly increased both BMSC proliferation rate and ALP activity, while it decreased the indicators of oxidative stress, caspase-3 activity, and the apoptosis rate of BMSCs induced by H2O2.. Conclusion: PNS, acting as a biological antioxidant, had a protective effect on H2O2-induced apoptosis in cultured rabbit BMSCs by decreasing oxidative stress and down-regulating caspase-3. [Copyright &y& Elsevier]

Panax notoginseng (P. notoginseng) has excellent medicinal and food dual-use characteristics. However, P. notoginseng with a unique origin label has become the target of fraud because of people confusing or hiding its origin. In this study, an untargeted nuclear magnetic resonance (NMR)-based metabolomics approach was used to discriminate the geographical origins of P. notoginseng from four major producing areas in China. Fifty-two components, including various saccharides, amino acids, saponins, organic acids, and alcohols, were identified and quantified through the NMR spectrum, and the area-specific geographical identification components were further screened. P. notoginseng from Yunnan had strong hypoglycemic and cardiovascular protective effects due to its high acetic acid, dopamine, and serine content, while P. notoginseng from Sichuan was more beneficial for diseases of the nervous system because of its high content of fumarate. P. notoginseng from Guizhou and Tibet had high contents of malic acid, notoginsenoside R1, and amino acids. Our results can help to distinguish the geographical origin of P. notoginseng and are readily available for nutritional recommendations in human consumption. [ABSTRACT FROM AUTHOR]

The core rhizosphere microbiome is critical for plant fitness. However, its contribution to the belowground biomass and saponin contents of Panax notoginseng remains unclear. High-throughput sequencing of amplicon and metagenome was performed to obtain the microbiome profiles and functional traits in P. notoginseng rhizosphere across a large spatial scale. We obtained 639 bacterial and 310 fungal core OTUs, which were mainly affected by soil pH and organic matter (OM). The core taxa were grouped into four ecological clusters (i.e. high pH, low pH, high OM and low OM) for sharing similar habitat preferences. Furthermore, structural equation modelling (SEM) and correlation analyses revealed that the diversity and composition of core microbiomes, as well as the metagenome-derived microbial functions, were related to belowground biomass and saponin contents. Key microbial genera related to the two plant indicators were also identified. In short, this study explored the main driving environmental factors of core microbiomes in the P. notoginseng rhizosphere and revealed that the core microbiomes and microbial functions potentially contributed to the belowground biomass and saponin contents of the plant. This work may enhance our understanding of interactions between microbes and perennial plants and improve our ability to manage root microbiota for the sustainable production of herbal medicine., (© 2022 Applied Microbiology International and John Wiley & Sons Ltd.)

Panax notoginseng (P. notoginseng) is a famous traditional Chinese Medicine and is widely used for functional food in folk. Panax notoginseng saponins (PNS) were the main active components in P. notoginseng. The composition and number of the side chain glycosyl in PNS show great influence on the activity of ginsenosides and can be changed greatly by enzyme conversion. With the development of biotechnology in food processing industry, the anti-tumor potential of PNS deserves attention. In this study, enzyme transformed PNS products (E-PNS) were prepared by recombinant thermostable β-Glucosidase Tpebgl3 from PNS. Through the optimization of enzyme catalytic condition, 380 mg E-PNS was obtained from the transformation of 500 mg PNS by 8000 U Tpebgl3 at 85 °C in 40 ml water for 2 h. The main components in E-PNS were notoginsenoside R2, ginsenoside Rh1 and Rg3. Both in vitro and in vivo results showed that the anti-tumor activity of E-PNS was better than PNS significantly. The IC 50 value of E-PNS against 4T1 cells was 132 μg/ml. E-PNS could induce the cell apoptosis of 4T1 cells through death receptor and mitochondrial pathway in a dose and time-dependent manner. The phosphorylation of protein kinase B (AKT) in proliferation signaling pathway was inhibited by E-PNS. In tumor tissue, the pathological score decreased and angiogenesis were inhibited significantly by the treatment of E-PNS. The study suggested that Tpebgl3 has great potential and prospect in promoting the application of P. notoginseng as functional food or food addition in cancer prevention or treatment. [Display omitted] [ABSTRACT FROM AUTHOR]

Introduction: Panax notoginseng, a medicinal herb in China, is attacked by several pathogens during its cultivation. Dazomet (DZ) is a soil fumigant that is effective in controlling soil-borne pathogens, but its long-term effects on P. notoginseng growth and soil properties are unknown. Methods: We conducted field experiments over two consecutive years to assess the impact of three concentrations of DZ fumigation (35 kg/666.7 m2, 40 kg/666.7 m2, and 45 kg/666.7 m2) on soil physicochemical properties, microbial diversity, and P. notoginseng growth. Correlation analyses were performed between microbial community changes and soil properties, and functional predictions for soil microorganisms were conducted. Results: DZ fumigation increased total nitrogen, total phosphorus, total potassium, available phosphorus, available potassium, and ammonia nitrogen levels in the soil. DZ fumigation promoted the nutrient accumulation and improvement of agronomic traits of P. notoginseng, resulted in a 2.83-3.81X yield increase, with the highest total saponin content increasing by 24.06%. And the 40 kg/666.7 m2 treatment had the most favorable impact on P. notoginseng growth and saponin accumulation. After DZ fumigation, there was a decrease in the relative abundance of pathogenic fungi such as Fusarium, Plectosphaerella, and Ilyonectria, while beneficial bacteria such as Ramlibacter, Burkholderia, and Rhodanobacteria increased. The effects of fumigation on soil microorganisms and soil physicochemical properties persisted for 18 months post-fumigation. DZ fumigation enhanced the relative abundance of bacteria involved in the biosynthesis of secondary metabolites and arbuscular mycorrhizal fungi, reduced the relative abundance of plant-animal pathogenic fungi, reduced the occurrence of soil-borne diseases. Conclusion: In conclusion, DZ fumigation enhanced soil physicochemical properties, increased the proportion of beneficial bacteria in the soil, and rebalanced soil microorganism populations, consequently improving the growth environment of P. notoginseng and enhancing its growth, yield, and quality. This study offers a theoretical foundation for DZ fumigation as a potential solution to the continuous cropping issue in perennial medicinal plants such as P. notoginseng. [ABSTRACT FROM AUTHOR]

The simultaneous identification and quantification of target metabolites from herbal medicines are difficult to implement by the full-scan MS based nontargeted metabolomics approaches. Here an in-source multiple collision-neutral loss filtering (IMC-NLF) based nontargeted metabolomics approach is developed and applied to identify and quantify the variations of malonyl-ginsenosides, a common group of acyl saponins with potential anti-diabetic activity, among Panax ginseng , P. quinquefolius , and P. notoginseng . The key steps of the IMC-NLF strategy are the acquisition of specific high-resolution neutral loss data and the efficient filtering of target precursor ions from the full-scan spectra. Using a hybrid LTQ-Orbitrap mass spectrometer after UHPLC separation, abundant in-source product ions, [M-H-CO 2 ] - (due to the vulnerability of the carboxyl group) and [M-H-Mal.] - , were generated at the energies of 70 V and 90 V, respectively. After spectral deconvolution, the generated peak list was screened by dual NLF using a Neutral Loss MS Finder software (NL of 43.9898 Da for CO 2 and 86.0004 Da for the malonyl substituent). By combining the precursor ions list-triggered HCD-MS/MS and basic hydrolysis, a total of 101 malonyl-ginsenosides (including 69 from P. ginseng , 52 from P. quinquefolius , and 44 from P. notoginseng ) were identified or tentatively characterized. The variations of 81 characterized malonyl-ginsenosides among 45 batches of Ginseng samples were statistically analyzed disclosing ten potential markers. It is the first systematic analysis of malonyl-ginsenosides. The IMC-NLF approach by a single analytical platform is promising in targeted analyses of modification-specific metabolites in metabolomics and drug metabolism. [ABSTRACT FROM AUTHOR]

This study investigated the effect of nitrogen (N) and potassium (K) formula fertilization on cadmium (Cd) accumulation in P. notoginseng. Field investigations as well as formulated N and K fertilizers application experiments were conducted. Field investigations showed that Cd accumulation decreased PNS content in the main roots of P. notoginseng, while PNS content was promoted by soil available potassium (AK) and K in the main roots. The Cd content in P. notoginseng and the bioavailable Cd content in the soil decreased with the increasing of total K (TK) and AK in the soil. The increase of soil pH, total organic matter (TOM) and cation exchange capacity (CEC) values can reduce the bioavailable Cd content in soil, thus reducing the Cd accumulation in P. notoginseng. Under current fertilization in P. notoginseng cultivation, decreased N fertilization can alleviate the deterioration of soil physical and chemical properties. Under identical N fertilization, increasing K fertilization promoted the PNS accumulation (0.3–38.3%), also improved soil physical and chemical properties. Formulated N and K fertilizers application (1:2) experiments showed that reducing application of N and increasing K fertilization could reduce the bioavailable Cd content in soil, and the Cd content also decreased by 0.5–69.6% in P. notoginseng. Abbreviation: PNS: P. notoginseng saponins; F(EXC): Exchangeable fraction; F(Carb): Bound to carbonates fraction; F(Fe-MnOX): Bound to iron and manganese oxides fraction; F(OM): Bound to organic matter fraction; F(RES): Residual fraction; AK: Available potassium; TK: Total potassium; CEC: Cation exchange capacity; TOM: Total organic matter [ABSTRACT FROM AUTHOR]

Panax notoginseng saponins (PNSs) are used as industrial raw materials to produce many drugs to treat cardio-cerebrovascular diseases. However, it is a heat-sensitive plant, and its large-scale artificial cultivation is impeded by high temperature stress, leading to decreases in productivity and PNSs yield. Here, we examined exogenous foliar leucine to alleviate heat stress and explored the underlying mechanism using metabolomics. The results indicated that 3 and 5 mM exogenous foliar leucine significantly alleviated heat stress in one-year- and two-year-old P. notoginseng in pots and field trials. Exogenous foliar leucine enhanced the antioxidant capacity by increasing the activities of antioxidant enzymes (POD, SOD) and the contents of antioxidant metabolites (amino acids). Moreover, exogenous foliar leucine enhanced carbohydrate metabolism, including sugars (sucrose, maltose) and TCA cycle metabolites (citric acid, aconitic acid, succinic acid and fumaric acid), in P. notoginseng leaves, stems, and fibrous roots to improve the energy supply of plants and further alleviate heat stress. Field experiments further verified that exogenous foliar leucine increased the productivity and PNSs accumulation in P. notoginseng. These results suggest that leucine application is beneficial for improving the growth and quality of P. notoginseng under heat stress. It is therefore possible to develop plant growth regulators based on leucine to improve the heat resistance of P. notoginseng and other crops. [ABSTRACT FROM AUTHOR]

Under different concentrations of cadmium treatments (Cd2+ concentration: 0, 0.6, 6.0, 12.0 mg/kg, prepared with CdCl2•2.5H2O), pot experiments were conducted to study the application of two calcium-containing substances (Ca2+ concentration: 0, 180, 360 mg/kg, prepared with calcium chloride CaCl2 and lime Ca(OH)2) on the yields of flavonoids and saponins of P. notoginseng. The results showed that the application of calcium chloride and lime could increase the biomass of the main root of P. notoginseng and significantly reduce the cadmium contents. Under 0 and 0.6 mg/kg cadmium treatment, the cadmium content in the main root of P. notoginseng did not exceed the standard value (Cd≤0.5 mg/kg1, GB/T 19086–2008). Under 0.6 mg/kg cadmium treatment, the application of calcium chloride increased the yield of flavonoids in the main root of P. notoginseng. The yield of flavonoids in the main root of P. notoginseng decreased under the treatment of 360 mg/kg Ca2+ lime and 0.6 mg/kg Cd2+. Under the treatments of 6.0 mg/kg and 12.0 mg/kg cadmium stress, calcium chloride and lime reduced the yield of flavonoids in P. notoginseng by 14.2%-35.2%. There was a significant positive logarithmic function correlation between flavonoid yield and ratio of Ca/Cd treatment concentration (P < 0.05). Under the treatment of 0.6 mg/kg Cd2+ with 180 mg/kg Ca2+ (calcium chloride), yields of Rg1, Rb1 and total saponins of P. notoginseng reached the highest and were increased by 114.9%, 323.0% and 172.4%, respectively, compared with no calcium chloride treatment. Under different Cd treatment levels and lime application, the yields of Rg1, Rb1 and total saponins of P. notoginseng decreased and the yield of R1 remained relatively stable. Under a certain concentration of cadmium treatment, appropriate calcium chloride and lime application could increase or maintain the biomass, saponin and flavonoid yields of the main root of P. notoginseng. Yields of R1, Rg1, Rb1 and SPN were influenced due to interaction of Cd2+ and Ca2+. In order to ensure the cadmium content of P. notoginseng less than the standard value and optimal yields of flavonoids and saponins, it could be recommended to apply calcium chloride or lime 1125 kg/hm (180 mg/kg Ca2+) under 0.6 mg/kg Cd2+ treatment. [ABSTRACT FROM AUTHOR]

Nitrogen (N) is an important macronutrient and is comprehensively involved in the synthesis of secondary metabolites. However, the interaction between N supply and crop yield and the accumulation of effective constituents in an N-sensitive medicinal plant Panax notoginseng (Burkill) F. H. Chen is not completely known. Morphological traits, N use and allocation, photosynthetic capacity and saponins accumulation were evaluated in two- and three-year-old P. notoginseng grown under different N regimes. The number and length of fibrous root, total root length and root volume were reduced with the increase of N supply. The accumulation of leaf and stem biomass (above-ground) were enhanced with increasing N supply, and LN-grown plants had the lowest root biomass. Above-ground biomass was closely correlated with N content, and the relationship between root biomass and N content was negatives in P. notoginseng (r = −0.92). N use efficiency-related parameters, NUE (N use efficiency, etc.), Nc (N content in carboxylation system component) and Pn (the net photosynthetic rate) were reduced in HN-grown P. notoginseng. SLN (specific leaf N), Chl (chlorophyll), NL (N content in light capture component) increased with an increase in N application. Interestingly, root biomass was positively correlated with NUE, yield and Pn. Above-ground biomass was close negatively correlated with photosynthetic N use efficiency (PNUE). Saponins content was positively correlated with NUE and Pn. Additionally, HN improved the root yield of per plant compared with LN, but reduced the accumulation of saponins, and the lowest yield of saponins per unit area (35.71 kg·hm−2 ) was recorded in HN-grown plants. HN-grown medicinal plants could inhibit the accumulation of root biomass by reducing N use and photosynthetic capacity, and HN-induced decrease in the accumulation of saponins (C-containing metabolites) might be closely related to the decline in N efficiency and photosynthetic capacity. Overall, N excess reduces the yield of root and C-containing secondary metabolites (active ingredient) in N-sensitive medicinal species such as P. notoginseng [ABSTRACT FROM AUTHOR]

Panax notoginseng is a highly valued perennial medicinal herb plant in Yunnan Province, China, and the taproots are the main medicinal parts that are rich in active substances of P. notoginseng saponins. The main purpose of this study is to uncover the physiological and molecular mechanism of Panax notoginseng saponin accumulation triggered by methyl jasmonate (MeJA) under arbuscular mycorrhizal fungi (AMF) by determining physiological indices, high-throughput sequencing and correlation analysis. Physiological results showed that the biomass and saponin contents of P. notoginseng, the concentrations of jasmonic acids (JAs) and the key enzyme activities involved in notoginsenoside biosynthesis significantly increased under AMF or MeJA, but the interactive treatment of AMF and MeJA weakened the effect of AMF, suggesting that a high concentration of endogenous JA have inhibitory effect. Transcriptome sequencing results indicated that differential expressed genes (DEGs) involved in notoginsenoside and JA biosynthesis were significantly enriched in response to AMF induction, e.g., upregulated genes of diphosphocytidyl-2-C-methyl-derythritol kinases (ISPEs), cytochrome P450 monooxygenases (CYP450s)_and glycosyltransferases (GTs), while treatments AMF-MeJA and salicylhydroxamic acid (SHAM) decreased the abundance of these DEGs. Interestingly, a high correlation presented between any two of saponin contents, key enzyme activities and expression levels of DEGs. Taken together, the inoculation of AMF can improve the growth and saponin accumulation of P. notoginseng by strengthening the activities of key enzymes and the expression levels of encoding genes, in which the JA regulatory pathway is a key link. This study provides references for implementing ecological planting of P. notoginseng, improving saponin accumulation and illustrating the biosynthesis mechanism. [ABSTRACT FROM AUTHOR]

Keywords: Panax notoginseng; genome; triterpenoid saponins; dencichine EN Panax notoginseng genome triterpenoid saponins dencichine 869 871 3 05/20/21 20210501 NES 210501 I Panax notoginseng i (2n = 2x = 24), reputed as a valuable medicinal plant, belongs to the Araliaceae family (Figure 1a). In conclusion, this high-quality genome assembly of I P. notoginseng i provides novel insights into unique saponins distribution pattern and reveals possible dencichine biosynthetic pathway. The chromosome-scale high-quality genome assembly of Panax notoginseng provides insight into dencichine biosynthesis Gene family analysis of I P. notoginseng i and 11 other angiosperms suggest I P. notoginseng i genes were clustered into 17,306 families and I P. notoginseng i had much less multiple-copy orthologs compared with I P. ginseng i (Figure 1f). [Extracted from the article]

Cadmium (Cd) pollution poses potential safety risks for Panax notoginseng cultivation, a medicinal plant in Yunnan. Under exogenous Cd stress, field experiments were conducted to understand the effects of lime (0, 750, 2250 and 3750 kg hm−2) applied and oxalic acid (0, 0.1 and 0.2 mol L−1) leaves sprayed on Cd accumulation, antioxidant system and medicinal components of P. notoginseng. The results showed that Lime and foliar spray of oxalic acid were able to elevate Ca2+ and alleviate Cd2+ toxicity in P. notoginseng under Cd stress. The addition of lime and oxalic acid increased the activities of antioxidant enzymes and alters osmoregulator metabolism. The most significant increase in CAT activities increased by 2.77 folds. And the highest increase of SOD activities was 1.78 folds under the application of oxalic acid. While MDA content decreased by 58.38%. There were very significant correlation with soluble sugar, free amino acid, proline and soluble protein. Lime and oxalic acid were able to increase calcium ions (Ca2+), decrease Cd content and improve the stress resistance of P. notoginseng, while increasing the production of total saponins and flavonoids. Cd content were the lowest, 68.57% lower than controls, and met the standard value (Cd ≤ 0.5 mg kg−1, GB/T 19086-2008). The proportion of SPN was 7.73%, which reached the highest level of all treatments, the flavonoids content increased significantly by 21.74%, which reached the medicinal standard value and optimal yield. [ABSTRACT FROM AUTHOR]

The root of Panax notoginseng (Burk.), in which saponins are the major active components, is a famous traditional Chinese medicine used to stop bleeding and to decrease inflammation and heart disease. Inflorescence removal increases the yield and quality of P. notoginseng , but the underlying molecular mechanisms are unknown. Here, the differences between inflorescence-removal treatment and control groups of P. notoginseng were compared using transcriptomics and metabolomics analyses. Illumina sequencing of cDNA libraries prepared from the rhizomes, leaves and roots of the two groups independently identified 6,464, 4,584, and 7,220 differentially expressed genes (DEG), respectively. In total, 345 differentially expressed transcription factors (TFs), including MYB and WRKY family members, were induced by the inflorescence-removal treatment. Additionally, 215 DEGs involved in saponin terpenoid backbone biosynthetic pathways were identified. Most genes involved in the mevalonic acid (MVA) and methylerythritol phosphate (MEP) pathways were activated by inflorescence removal. The co-expression analysis showed that the low expression levels of flavonoid biosynthesis-related genes (e.g., C4H and F3H) decreased the biosynthesis and accumulation of some flavonoids after inflorescence removal. The results not only provide new insights into the fundamental mechanisms underlying the poorly studied inflorescence-removal process in P. notoginseng and other rhizome crops, but they also represent an important resource for future research on gene functions during inflorescence-removal treatments and the reproductive stage. [ABSTRACT FROM AUTHOR]

Excessive nitrogen (N) application and potassium (K) supplement deficiency is a common problem in Panax notoginseng cultivation. However, synergistic effects of lowering N and increasing K application on yield and quality of P. notoginseng have not been reported. Field experiments in two locations with different N and K combined application were conducted to study the effects on yield and quality. Then, the saponin accumulation mechanisms were explored by pot and hydroponic culture with 2- or 3-year-old seedlings. The investigation showed that 70% of P. notoginseng cultivation fields reached abundant levels of total nitrogen (TN) but had deficient levels of total potassium (TK), which may be detrimental to balance the N/K uptake of P. notoginseng. Moreover, the average biomass was 18.9 g, and P. notoginseng saponin (PNS) content was 6.95%; both were influenced by the N/K values of P. notoginseng. The field experiments indicated that compared to the conventional N and K application (N:K = 2:1), lowering N and increasing K application (N:K = 1:2) decreased root rot rate by 36.4–46.1% and increased survival rate, root biomass, and yield, as well as PNS content by 17.9–18.3, 5.7–32.9, 27.8–57.1, and 5–10%, respectively. The mechanism of lowering N and increasing K application on the PNS content improving was due to the decreasing of N/K value, which promoted photosynthesis, sugar accumulation, and the expression of saponin biosynthesis genes. Therefore, lowering N and increasing K application to the ratio of 1:2 would have great potential to improve the synergistic effect on yield and quality of P. notoginseng cultivation. [ABSTRACT FROM AUTHOR]

Abstract: Panax, a genus of the Araliaceae family, is an important herbal group in traditional Chinese medicine (TCM). Nine species and three varieties are included in the genus of Panax, in which nearly all species have been used for medicinal purposes. Among them, Panax notoginseng (Burk) F. H. Chen, Panax ginseng C. A. Meyer and Panax quinquefolius L. are the most representative and valuable herbs world-wide, with a long history of cultivation. As the main bioactive chemical constituents, saponins with different aglycones are the major components in various Panax spp., and their pharmacological activities are mainly reflected in the effects on blood system, cardio- and cerebro-vascular systems, nervous system, metabolism, and immune regulation. Researchers of Kunming Institute of Botany (KIB), Chinese Academy of Sciences (CAS), have put many efforts into conducting the investigations on Panax species. Herein, we reviewed the research progress on Panax spp. in KIB, CAS, over the past few decades, from the aspects of history and origin, phytochemistry and pharmacological activities.Graphical Abstract: [ABSTRACT FROM AUTHOR]

Abstract: Triterpene saponins are main active constituents of Panax notoginseng. Metabolites profiling of 12 triterpene saponins was analyzed by high‐performance liquid chromatography–mass spectrometry in leaf, petiole, and root extracts of P. notoginseng. Most of the 20(S)‐protopanaxatriol (PPT) type saponins, except ginsenoside Re, were mainly distributed in roots, while 20(S)‐protopanaxadiol (PPD) type saponins were detected among various tissues. The total content of PPD‐type saponins decreased in the order of leaf, petiole, and root. The expression patterns of four key genes (PnFPS, PnSQS, PnDS, and PnSE) in the triterpene saponin biosynthetic pathway were measured by real‐time quantitative PCR (RT‐qPCR). All the four investigated genes showed high expression levels in leaf. A gene‐to‐metabolite network was constructed through canonical correlation analysis. The results indicated that the expression levels of PnFPS, PnSQS, PnDS, and PnSE had high correlation with PPD‐type saponins ginsenoside Rb2, Rb3, and Rc, while PnSQS was also highly correlated with Rb1. Combining metabolic profiling, RT‐qPCR, and gene‐to‐metabolite network, we inferred that the leaf of P. notoginseng was the main biosynthesis site of PPD‐type saponins Rb2, Rb3, and Rc. The contribution to the biosynthesis of ginsenosides Rb2, Rb3, and Rc was in the order of PnSE > PnDS > PnSQS > PnFPS. PnSE and PnDS should be the preferred targets to regulate the production of PPD‐type saponins Rb2, Rb3, and Rc in P. notoginseng by plant metabolic engineering. [ABSTRACT FROM AUTHOR]

Essential oils (EOs) have been proposed as an alternative to conventional pesticides to inhibit fungal pathogens. However, the application of EOs is considerably limited due to their highly volatile nature and unpredictable effects on other microbes. In our study, the composition of bacterial and fungal communities from the rhizosphere soil of P. notoginseng under four treatment levels of Alpinia officinarum Hance EO was characterized over several growth stages. Leaf weight varied dramatically among the four EO treatment levels after four months of growth, and the disease index at a low concentration (0.14 mg/g) of EO addition was the lowest among the P. notoginseng growth stages. The content of monomeric saponins was elevated when EO was added. Bacterial and fungal diversity in the absence of plants showed a decreasing trend with increasing levels of EO. Bacterial diversity recovery was more correlated with plant growth than was fungal diversity recovery. Compared with the control (no EO addition), a low concentration of EO significantly accumulated Actinomycota, including Acidothermus, Blastococcus, Catenulispora, Conexibacter, Rhodococcus, and Sinomonas, after one month of plant-microbial interaction. Overall, the results showed that both the plant growth stage and EOs drive changes in the microbial community composition in the rhizosphere of P. notoginseng. Plant development status had a stronger influence on bacterial diversity than on fungal diversity. EO had a more significant effect on fungal community composition, increasing the dominance of Ascomycota when EO concentration was increased. Under the interaction of P. notoginseng growth and EO, a large number of bacterial genera that have been described as plant growth-promoting rhizobacteria (PGPR) responded positively to low concentrations of EO application, suggesting that EO may recruit beneficial microbes in the root zone to cope with pathogens and reduce root rot disease. These results offer novel insights into the relationship between EO application, altered microbial communities in the plant roots, plant growth stage, and disease occurrence. [ABSTRACT FROM AUTHOR]

Endophytes in plants may be co-producer of the bioactive compounds of their hosts. We conducted a study to bioprospect for saponin-producing endophytic fungi from Panax notoginseng and evaluate the antimicrobial activity of saponins. Two novel fungal endophytes, Fusarium sp. PN8 and Aspergillus sp. PN17, were isolated from traditional Chinese medicinal herb P. notoginseng. After eight days of fermentation, the total saponins produced in the culture broth of PN8 and PN17 were 1.061 and 0.583 mg mL -1 , respectively. The saponin extracts exhibited moderate to high (inhibition zone diameter 15.7-28.4 mm, MIC 1.6-12.5 mg mL -1 ) antimicrobial activity against pathogens tested. Further analysis showed that triterpenoid saponins produced by Fusarium PN8 were Rb 1 , Rd and 20(S)-Rg 3 , while Aspergillus PN17 had the ability to synthesise ginsenoside Re, Rd and 20(S)-Rg 3 . The isolated endophytes may be used as potential sources for microbial production of plant secondary metabolites and for antimicrobial agents.

Panax notoginseng is an important Chinese medicinal plant. Saponins are the major bioactive secondary metabolites with a wide range of medicinal and commercial value in P. notoginseng, so it is crucial to develop environmentally friendly methods to increase their production. The symbiotic relationship between endophytic bacteria and host plants offers a sustainable approach to enhance secondary metabolite biosynthesis. In this study, it was reported that the co-cultivation of an endophytic bacterium Enterobacter cloacae PN7, isolated from P. notoginseng and its host plant could greatly promote saponin accumulation in the root of seedlings. After six days of PN7 treatment, the total saponin concentration reached 21.64 mg/g, representing a 2.01-fold increase over the control. Transcriptome sequencing revealed that PN7 induction upregulated key genes in the saponin biosynthetic pathway (including DXS, HMGR, PMK, DS, CYP450, and GTs), modulated 253 plant hormone signaling genes (such as those related to JA, ETH, and ABA), and affected 284 transcription factor genes and 47 ABC transporter genes. Co-expression network analysis identified DEGs related to plant hormone signaling, transcription factors, and ABC transporters in saponin biosynthesis and distribution. The results suggested that JA signaling, mediated by transcription factors, such as bHLH and MYBs, and its interaction with ETH, played crucial roles in saponin biosynthesis. Additionally, potential ABC transporter candidates involved in saponin transport were identified. This study highlights the role of endophytic bacteria in enhancing saponin production in P. notoginseng and opens avenues for further research on microbial-plant interactions in secondary metabolite production., Competing Interests: Declarations. Conflict of interest: The authors declare no competing interests. Ethical Approval and Consent to Participate: This research study has no connection to human participants or animals by any of the authors. Consent for Publication: All authors read and agreed to the submission of this research work to this journal., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Sanqi, the root of Panax notoginseng, has long been recognized for its therapeutic effects on cardiovascular diseases. Saponins, including ginsenosides and notoginsenosides, are the main bioactive components of P. notoginseng. The biosynthesis of saponins is closely related to the defense responses orchestrated by endogenous hormones., Results: To provide new insights into the underlying role of phytohormone jasmonic acid (JA) in the synthesis and regulation of saponins, we performed an ultra-performance liquid chromatography analysis of different tissues of P. notoginseng aged 2-4 years. Moreover, by combined evaluation of saponin content and transcriptome profiling of each tissue, the spatial and temporal distribution of saponins was analyzed. N notoginsenoside R1, ginsenoside Rb1 and ginsenoside Rd accumulated in the underground tissues, including the root, tuqi, fibril and rhizome. In agreement with this data, the corresponding genes of the endogenous hormone JAs, especially coronatine insensitive 1 (COI1) and myelocytomatosis proteins 2 (MYC2), were predominantly expressed in the underground tissues. The tissue- and age-specific distribution of saponins was consistent with the expression of genes involved in JA biosynthetic, metabolic and signaling pathways., Conclusion: The present study has revealed the temporal and spatial effects of endogenous phtohormones in the synthesis and regulation of notoginsenosides, which will provide a significant impact on improving the ecological planting technology, cultivating new high-quality varieties and protecting the rare resources of medicinal P. notoginseng. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Main conclusion: Transcriptome analysis revealed high expression of saponin biosynthetic genes may account for highly accumulated saponins in 3-year-oldPanax notoginsengroots andDSandCYP716A47-likewere functionally verified by transgenic tobacco.Panax notoginseng is a well-known traditional medical herb that contains bioactive compounds known as saponins. Three major dammarene-type triterpene saponins including R1, Rb1, and Rg1 were found to be highly accumulated in the roots of 3-year-old plants when compared to those of 1-year-old plants. However, the underlying cellular mechanism is poorly understood. In this study, transcriptome analysis revealed that most genes involved in saponin biosynthesis in P. notoginseng roots augmented during their growth periods. The analysis of the KEGG pathway indicated that the primary metabolism, cell growth, and differentiation were less active in the roots of 3-year-old plant; however, secondary metabolisms were enhanced, thus providing molecular evidence for the harvesting of P. notoginseng roots in the 3rd year of growth. Furthermore, the functional role of DS and CYP716A47-like, two of the candidate genes involved in saponin biosynthesis isolated from P. notoginseng, were verified via overexpression in cultivated tobacco. Approximately, 0.325 µg g−1 of dammarenediol-II and 0.320 µg g−1 of protopanaxadiol were recorded in the dry leaves of transgenic tobacco overexpressed with DS and both DS and CYP716A47-like, respectively. This study provides insights into the molecular mechanisms for saponin accumulation in P. notoginseng roots during its growth period and paves a promising way to produce dammarenediol-II and protopanaxadiol via transgenic techniques. [ABSTRACT FROM AUTHOR]

Background: MicroRNAs (miRNAs) are small, non-coding RNAs that are important regulators of gene expression, and play major roles in plant development and their response to the environment. Root extracts from Panax notoginseng contain triterpene saponins as their principal bioactive constituent, and demonstrate medicinal properties. To investigate the novel and conserved miRNAs in P. notoginseng, three small RNA libraries constructed from 1-, 2-, and 3-year-old roots in which root saponin levels vary underwent high-throughput sequencing. Methods: P. notoginseng roots, purified from 1-, 2-, and 3-year-old roots, were extracted for RNA, respectively. Three small libraries were constructed and subjected to next generation sequencing. Results: Sequencing of the three libraries generated 67,217,124 clean reads from P. notoginseng roots. A total of 316 conserved miRNAs (belonging to 67 miRNA families and one unclassified family) and 52 novel miRNAs were identified. MIR156 and MIR166 were the largest miRNA families, while miR156i and miR156g showed the highest abundance of miRNA species. Potential miRNA target genes were predicted and annotated using Cluster of Orthologous Groups, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes. Comparing these miRNAs between root samples revealed 33 that were differentially expressed between 2- and 1-year-old roots (8 increased, 25 decreased), 27 differentially expressed between 3- and 1-year-old roots (7 increased, 20 decreased), and 29 differentially expressed between 3- and 2-year-old roots (8 increased, 21 decreased). Two significantly differentially expressed miRNAs and four miRNAs predicted to target genes involved in the terpenoid backbone biosynthesis pathway were selected and validated by quantitative reverse transcription PCR. Furthermore, the expression patterns of these six miRNAs were analyzed in P. notoginseng roots, stems, and leaves at different developmental stages. Conclusions: This study identified a large number of P. notoginseng miRNAs and their target genes, functional annotations, and gene expression patterns. It provides the first known miRNA profiles of the P. notoginseng root development cycle. [ABSTRACT FROM AUTHOR]

Ginsenosides are the primary active substance in ginseng plants and have a variety of benefits. However, its light and heat stability are weak and easy to decompose. This study used gum arabic (GA) and maltodextrin (MD) as wall materials, and 1% Tween 80 was used as emulsifier. Response surface methodology was used to optimize the preparation process of total saponins in the stems-leaves of Panax notoginseng (SLPNs) (SSLP) microcapsules by spray drying and freeze drying techniques. Under optimal process conditions, the two microcapsules have better solubility and lower moisture content (MC). The color of spray-dried SSLP microcapsules was greener and bluer, and the color was brighter. In morphology, the spray-dried SSLP microcapsules were spherical with a slightly shrunk surface, whereas the freeze-dried ones were lamellar and porous. The two microcapsules have strong stability under different storage conditions and in vitro gastrointestinal digestion simulation. In addition, both microcapsules and free SSLP contained multiple ginsenosides. At the same time, both microcapsules had good free radical scavenging ability. These results indicate that the microencapsulation technology could improve the stability and bioavailability of SSLP, which is expected to provide a reference for the intensive processing of the SLPN. PRACTICAL APPLICATION: After microencapsulation, the stem and leaf extract of Panax notoginseng improved its stability and taste, which laid a foundation for making more nutritious and better tasting food of the stem and leaf of P. notoginseng., (© 2024 Institute of Food Technologists.)

Abstract: Panax notoginseng has the effect of stimulating circulation to end stasis. Our study was designed to evaluate the anti-thrombotic effect of protoparaxotriol saponins (PTS) from P. notoginseng and the involved mechanisms. A thrombosis model was constructed, and the anti-thrombotic activity of PTS was determined by erythrocyte staining, heart rate, and blood flow velocity. In addition, quantitative real-time polymerase chain reaction was used to identify changes in the expression of genes related to coagulation, inflammation, and apoptosis. PTS alleviated arachidonic acid-induced caudal vein thrombosis, restored blood flow, and increased the area of cardiac erythrocyte staining, heart rate, and blood flow velocity. It reduced the ponatinib-induced cerebral thrombus area and decreased the intensity of erythrocyte staining. The quantitative polymerase chain reaction data showed that the anti-thrombotic effect of PTS was mediated by suppression of genes related to coagulation, inflammation, and apoptosis and also involved inhibition of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) pathways., Competing Interests: The authors report no conflicts of interest., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

Panax notoginseng (Burk.) F.H. Chen is a species of the Araliaceae family that inhabits southwestern China, Burma, and Nepal. It is cultivated on a commercial scale in Yunnan province, China, owing to its significance in traditional Chinese medicine. Panax notoginseng roots are usually yellow-white (HS); however, purple roots (ZS) have also been reported. The majority of P. notoginseng research has concentrated on the identification and production of natural chemicals in HS; however, there is little to no information about the composition of ZS. Using UPLC-MS/MS, we investigated the global metabolome profile of both ZS- and HS-type roots and discovered 834 metabolites from 11 chemical groups. There were 123 differentially accumulated metabolites (DAM) in the HS and ZS roots, which were classified as lipids and lipid-like molecules, polyketides, organoheterocyclic chemicals, and organooxygen compounds. We investigated the associated compounds in the DAMs because of the importance of anthocyanins in color and saponins and ginsenosides in health benefits. In general, we discovered that pigment compounds such as petunidin 3-glucoside, delphinidin 3-glucoside, and peonidin-3-O-beta-galactoside were more abundant in ZS. The saponin (eight compounds) and ginsenoside (26 compounds) content of the two varieties of roots differed as well. Transcriptome sequencing revealed that flavonoid and anthocyanin production genes were more abundant in ZS than in HS. Similarly, we found differences in gene expression in genes involved in terpenoid production and related pathways. Overall, these findings suggest that the purple roots of P. notoginseng contain varying amounts of ginsenosides and anthocyanins compared to roots with a creamy yellow color. [ABSTRACT FROM AUTHOR]

Panax notoginseng is a traditional Chinese medicine for the treatment of blood diseases, in which saponins were the main active components. Dammarenediol synthase (DS) is a key enzyme in the saponin synthesis pathway of P. notoginseng. The promoter is an important region to regulate gene expression, and the study of the promoter sequence provides important evidence for revealing the mechanism of gene expression regulation. However, there was still little research on the promoter function of P. notoginseng. In this study, the 1382 bp promoter upstream of DS from P. notoginseng was cloned and sequenced. The promoter sequence was analyzed by online databases. The plant expression vector fused with the β-glucuronidase gene was constructed and transferred into Agrobacterium tumefaciens. Then tobacco was injected, and its response to exogenous hormones (gibberellin and abscisic acid) was studied by transient expression to verify its unique action elements. The results showed that the tobacco leaves transferred with DS promoter had significantly increased GUS protease activity after spraying GA and ABA, indicating that both DS promoter can specifically and significantly respond to exogenous GA and ABA signal. These findings will help us to better understand the regulatory mechanisms of the upstream region of the DS gene and provide a basis for future research on the interaction of cis-acting elements of promoters with related transcription factors. [ABSTRACT FROM AUTHOR]

Saponins are kinds of antifungal compounds produced by Panax notoginseng to resist invasion by pathogens. Ilyonectria mors-panacis G3B was the dominant pathogen inducing root rot of P. notoginseng, and the abilities to detoxify saponins were the key to infect P. notoginseng successfully. To research the molecular mechanisms of detoxifying saponins in I. mors-panacis G3B, we used high-throughput RNA-Seq to identify 557 and 1519 differential expression genes (DEGs) in I. mors-panacis G3B with saponins treatments for 4H (Hours) and 12H (Hours) compared with no saponins treatments, respectively. Among these DEGs, we found 93 genes which were simultaneously highly expressed in I. mors-panacis G3B with saponins treatments for 4H and 12H, they mainly belong to genes encoding transporters, glycoside hydrolases, oxidation–reduction enzymes, transcription factors and so on. In addition, there were 21 putative PHI (Pathogen–Host Interaction) genes out of those 93 up-regulated genes. In this report, we analyzed virulence-associated genes in I. mors-panacis G3B which may be related to detoxifying saponins to infect P. notoginseng successfully. They provided an excellent starting point for in-depth study on pathogenicity of I. mors-panacis G3B and developed appropriate root rot disease management strategies in the future. [ABSTRACT FROM AUTHOR]