Panax ginseng as a perennial herb of Araliaceae, exhibits pharmacological effects such as central nervous system stimulation, anti-tumor properties, and cardiovascular and cerebrovascular protection. The B3 gene family plays a crucial role in growth and development, antioxidant activity, stress resistance, and secondary metabolism regulation of plants and has been extensively studied in various plants. However, the identification and analysis of the B3 gene family in P. ginseng have not been reported. In this study, a total of 145 B3 genes(PgB3s) with complete open reading frames(ORF) were identified from P. ginseng and classified into five subfamilies based on domain types. Through correlation analysis with ginsenoside content, SNP/InDels analysis, and interaction analysis with key enzyme genes, 15 PgB3 transcripts were found to be significantly correlated with ginsenoside content and exhibited a close interaction network with key enzyme genes involved in ginsenoside biosynthesis, which indicated that these genes may participate in the regulation of ginsenoside biosynthesis. Additionally, this study found that PgB3 genes exhibited induced expression in response to methyl jasmonate(MeJA) stress, which aligned with the presence of abundant stress response elements in their promoters, confirming the important role of the B3 gene family in P. ginseng in stress resistance. The results of this study revealed the potential functions of PgB3 genes in ginsenoside biosynthesis and stress response, providing a significant theoretical basis for further research on the functions of PgB3 genes and their regulatory mechanisms.
Panax/metabolism* ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism* ; Ginsenosides/biosynthesis* ; Multigene Family ; Phylogeny

The study is aimed to explore the effect of soil moisture content on ginsenoside biosynthesis and explain its mechanism from the perspectives of antioxidant enzyme system and gene expression of key enzymes in the pathway of ginsenoside synthesis. In the study,two years old Panax ginseng was used as the experimental material and three moisture gradient,40% of saturated water content( W1),60%( W2),80%( W3) were set up. The content of 11 monomeric saponins were determined by HPLC. With GAPDH as a reference gene,six key enzymes( HMGR,SS,β-AS,CYP716 A47,CYP716 A52 v2,CYP716 A53 v2) in ginseng saponin synthesis pathway expression were analyzed by fluorescent quantitative PCR and the activities of superoxide dismutase( SOD),peroxidase( POD),catalase( CAT) activity and MDA content were also determined. With the increase of soil water,the content of ginseng saponin and biomass showed an increasing trend. PPD( Rb1,Rc,Rb2,Rd,Rh2,Rb3,Rg3),PPT( Rg1,Re,Rf) ginsenoside,Ro and total ginsenoside reached the maximum value on August 30,were 9.92,5.48,0.63 mg·g-1,respectively. During the whole regulation period,the antioxidant activity of W3 was greater than that of W1,and the MDA content was less than that of W1. At W3,expression levels of β-AS,CYP716 A47 and CYP716 A53 v2 showed an increasing trend,while HMGR and SS genes showed relatively stable expression levels under various water conditions. According to the correlation analysis,HMGR and SS genes in the W3 treatment group were significantly positively correlated with PPD,PPT ginsenoside and Ro,CYP716 A52 v2 gene was significantly positively correlated with Ro,and CYP716 A47 gene was significantly positively correlated with PPD ginsenoside. There was a significant positive correlation between β-AS gene and PPD ginsenoside in W1 and W2 treatment. Therefore,W3 is the optimum moisture content,ginseng total saponins and monomer saponin content is the highest,the gene closely correlation with content of saponins and more conducive to the accumulation of ginsenosides.
Chromatography, High Pressure Liquid ; Ginsenosides ; biosynthesis ; Panax ; physiology ; Water ; physiology

The relationship between saponin content of in different parts of the organization and expression of ginsenoside biosynthesis related gene was obtained by the correlation analysis between saponin content and gene expression. The 14 tissue parts of were studied, six saponins in Samples (ginsenoside Rg₁, Re, Rb₁, Rc, Rb₂ and Rd), group saponins and total saponins were determined by high performance liquid chromatography and vanillin-sulfuric acid colorimetric method. Simultaneously, the expression levels of 7 ginsenoside biosynthesis related genes ( and ) in different tissues of were determined by Real-time fluorescence quantitative PCR. Although 7 kinds of ginsenoside biosynthesis related enzyme gene in the involved in ginsenoside synthesis, the expression of and P450 genes had no significant effect on the content of monosodium saponins, grouping saponins and total saponins, and had significant or extremely significant on the contents of single saponins Re, Rg1, Rb1, Rd, group saponin PPD and PPT, total saponin TMS and total saponin TS (<0.05 or <0.01). The biosynthesis of partial saponins, grouping saponins and total saponins in was affected by the interaction of multiple enzyme genes in the saponin synthesis pathway, the content of saponins in different tissues of was determined by the differences in the expression of key enzymes in the biosynthetic pathway. Therefore, this study further clarified that and was the key enzyme to control the synthesis of saponins in by correlation analysis, the biosynthesis of ginsenosides in was regulated by these five kind of enzymes in cluster co-expression of interaction mode.
Biosynthetic Pathways ; Chromatography, High Pressure Liquid ; Ginsenosides ; biosynthesis ; genetics ; Panax ; genetics ; Plant Roots ; Saponins ; biosynthesis ; genetics

Traditional herbal medicines, Panax ginseng, Panax quinquefolium and Panax notoginseng, attract our attention for their extensive and powerful pharmacological activities. Ginsenosides are the main active constituents of these medicinal herbs. The related glycosyltransferases involved in ginsenoside biosynthesis are the key enzymes which catalyze the last important step. Modification of ginsenoside aglycones by glycosyltransferases produces the complexity and diversity of ginsenosides, which have more extensive pharmacological activity. At present, ginsenoside aglycones and compound K have been obtained by synthetic biology. As the last step of ginsenoside biosynthesis, glycosylation of ginsenoside aglycones has been studied intensively in recent years. This review summarizes the basic strategies and research advances in studies on glycosyltransferases involved in ginsenoside biosynthesis, which is expected to lay the theoretical foundation for the in-depth research of biosynthetic pathway of ginsenosides and their production by synthetic biology.
Biosynthetic Pathways ; Ginsenosides ; biosynthesis ; Glycosyltransferases ; metabolism ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Synthetic Biology

3-Hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR), the first enzyme of mavalonic acid pathway, is one of the key devices involved in ginsenoside biosynthesis based on synthetic biology approach. The open reading frame of a novel HMGR gene from Panax ginseng (PgHMGR2) was cloned and analyzed in this study. PgHMGR2-encoding protein showed 71.6% sequence similarity to a P. ginseng HMGR in GenBank. The full-length cDNA sequence of PgHMGR2 containing 1 770 bp, which encodes 589 amino acids, was cloned by RT-PCR strategy from P. ginseng. The bioinformatic analysis showed that PgHMGR2-encoding protein contained two transmembrane regions and the HMG_CoA_reductase domain, without signal peptide. The protein sequence of PgHMGR2 had the highest sequence similarity (99%) with Panax quinquefolius HMGR (GenBank accession No. ACV65036). The expression level of PgHMGR2 was the highest in flower based on a real-time PCR analysis, followed by leaf and root, and the lowest was in stem. The result will provide a foundation for exploring the molecular function of PgHMGR2 involved in ginsenoside biosynthesis based on synthetic biology approach in P. ginseng plants.
Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Flowers ; metabolism ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Plant ; Ginsenosides ; biosynthesis ; Hydroxymethylglutaryl CoA Reductases ; genetics ; metabolism ; Open Reading Frames ; Panax ; genetics ; metabolism ; Phylogeny ; Plant Leaves ; metabolism ; Plant Roots ; metabolism ; Plant Stems ; metabolism ; Plants, Medicinal ; genetics ; metabolism ; Protein Structure, Secondary ; Sequence Homology, Amino Acid ; Synthetic Biology

Ginsenosides are the main active components of medicinal herbs including Panax ginseng and Panax quinquefolium, which have potent effects of anti-tumor, anti-inflammatory, antioxidant and apoptosis inhibition. But the low content of ginsenosides limits its development and usage. At present, how to improve the production of ginsenosides by biological technology has been a new research focus. Some advances in the biosynthesis of ginsenosides by tissue culture and biotransformation have been made in recent years. So far at least twenty genes related to the biosynthesis of ginsenosides from Panax genus plants have been cloned and functionally identified, which has laid a good foundation for the study on the synthetic biology of ginsenosides. This review outlines recent advances in several aspects and is expected to provide a theoretical support to the thorough research of the pathway and regulation of ginsenosides biosynthesis.
Bacteria ; metabolism ; Biotransformation ; Fungi ; metabolism ; Ginsenosides ; biosynthesis ; metabolism ; Panax ; growth & development ; Plants, Medicinal ; growth & development ; Synthetic Biology ; Tissue Culture Techniques

The authors reviewed the new technologies used for Panax genus research, including molecular identification technologies (especially for DNA barcoding), modern biotechnologies (e. g. the first generation and second generation sequencing technologies), and gene cloning and identification in this paper. These technologies have been successfully applied to species identification, transcriptome analysis, secondary metabolite biosynthetic pathway and the key enzyme function identification, indicating that the application of modern biotechnologies provide guarantee for the molecular identification of Panax genus. The application of modern biotechnologies also reveals the genetic information of transcriptome and functional genomics, and promotes the design of Panax plants genomic map. In summary, the application of the new technologies lay the foundation for clarifying the molecular mechanisms of ginsenoside biosynthesis and enforcing the in vitro synthesis of important natural products and new drugs in future.
Biotechnology ; methods ; Cloning, Molecular ; DNA Fingerprinting ; Ginsenosides ; biosynthesis ; Panax ; enzymology ; genetics ; metabolism ; Research Design

OBJECTIVETo establish the induction method and culture system of hairy roots of Panax japonicus, and determine ginsenoside Re contents.

METHODHairy roots of P. japonicus was obtained through infecting pre-incubated terrestrial stem with the Agrobacterium tumefaciens strain C58C1; its enlarging culture was carried out on the 1/2MS medium, and growing characteristics were measured. The transformation of T-DNA was examined by PCR and ginsenoside Re content was determined by HPLC.

RESULTA. tumefaciens strain C58C1 could make terrestrial stem of P. japonicus bring about hairy roots, the max inductivity was 90% when infecting for 25 min. The PCR examination result showed that rolB genes could be inserted into the hair roots of P. japonicus. All hairy roots dould synthesize ginsenoside Re, among them, the max content was PJ8 with 60. 26 mg x g(-1).

CONCLUSIONIt was reported for the first time that the induction method and culture system of hairy roots of P. japonicus were established successfully, which provided a foundation for producing high content ginsenoside Re through culturing the hairy roots of P. japonicus.

Agrobacterium tumefaciens ; genetics ; physiology ; Cells, Cultured ; Culture Techniques ; methods ; Ginsenosides ; biosynthesis ; Panax ; genetics ; growth & development ; metabolism ; microbiology ; Plant Roots ; genetics ; growth & development ; metabolism ; microbiology

OBJECTIVETo study whether ginsenoside (GS) can regulate the glucocorticoid receptor (GR) in mice with ischemic liver damage, and to preliminarily observe its dose-effect relationship for providing an experimental bases in seeking a new way to relieve the damage from view of GR.

METHODSAdult male SD mouse was used to establish liver ischemia model, and different doses (100, 50, and 25 mg/kg) of GS was given via gastric infusion before modeling. The maximal GR binding capacity (Bmax) of liver and the level of GR mRNA expression in liver were dynamically determined at various time points (2 h, 6 h, 12 h and 24 h) after modeling.

RESULTSCompared with the normal control group, GR Bmax and GR mRNA expression in model rats were lower obviously (P < 0.01). As compared with the control group, GR Bmax and GR mRNA expression in model rats treated with 50 mg/kg GS significantly raised at 2 h, 6 h, 12 h (P < 0.01), while the changes in modeling rats treated with other two doses of GS were of no statistical significance.

CONCLUSIONGS in dose of 50 mg/kg can elevate the GR Bmax of liver and the level of GR mRNA expression in liver of rats with ischemic damage.

Animals ; Gene Expression Regulation ; drug effects ; Ginsenosides ; pharmacology ; Ischemia ; physiopathology ; Liver ; blood supply ; drug effects ; metabolism ; Male ; Mice ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Receptors, Glucocorticoid ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors

OBJECTIVETo investigate the role of P38 signaling pathway in modulating the expression of cyclooxygenase-2 (COX-2) in the substantia nigra (SN) of mice with 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-induced Parkinson disease (PD), and explore the possible mechanism of the dopaminergic (DA) neuron death in PD and the effects of ginsenoside Rg1 on the P38 signaling pathway and DA neurons.

METHODSC57BL6 mice were treated with MPTP to produce the subacute PD model, and the behavioral changes were observed. Immunohistochemistry and Western blotting for tyrosine hydroxylase (TH), COX-2, prostaglandin E2 (PGE2) and phosphorylated P38 (p-P38) were used to observe the changes of positive cell number in the midbrain after treatment with ginsenoside Rg1.

RESULTSCompared with the control mice, the mice with PD presented with typical symptoms of PD. The number of p-P38-, COX-2-, and PGE2-positive cells significantly increased in the SN area 6 h after the 3rd injection of 30 mg/kg MPTP (P<0.01). The number of TH-positive neurons in the PD model group was substantially reduced by about 60% (P<0.01) in 24 h after the 5th injection of MPTP. In mice with ginsenoside Rg1 treatment, the number of p-P38-, COX-2-, and PGE2-positive cells was reduced obviously 6 h after the 3rd injection of MPTP as compared with that in the model group (P<0.01). The number of TH-positive neurons in the SN was decreased by only 30% (P<0.01 vs control group) 24h after the 5th injection of MPTP.

CONCLUSIONP38 signaling pathway may play an important role in modulating COX-2 expression in the SN in the early stage of MPTP-induced subacute PD, and ginsenoside Rg1 may act on the P38 signaling pathway to protect the DA neurons in PD.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine ; Animals ; Blotting, Western ; Cyclooxygenase 2 ; biosynthesis ; Ginsenosides ; pharmacology ; Immunohistochemistry ; Male ; Mice ; Mice, Inbred C57BL ; Neurons ; drug effects ; metabolism ; pathology ; Parkinson Disease, Secondary ; chemically induced ; metabolism ; physiopathology ; Signal Transduction ; drug effects ; Substantia Nigra ; drug effects ; metabolism ; pathology ; p38 Mitogen-Activated Protein Kinases ; metabolism