Transcriptome-Wide Identification and Functional Analysis of PgSQE08-01 Gene in Ginsenoside Biosynthesis in Panax ginseng C.A.Mey.

Panax ginseng C.A.Mey.is an important plant species used in traditional Chinese medicine,whose primary active ingredient is a ginsenoside.Ginsenoside biosynthesis is not only regulated by transcription factors but also controlled by a variety of structural genes.Nonetheless,the molecular mechanism underlying ginsenoside biosynthesis has always been a topic in the discussion of ginseng secondary metabolites.Squalene epoxidase(SQE)is a key enzyme in the mevalonic acid pathway,which affects the biosynthesis of secondary metabolites such as terpenoid.Using ginseng transcriptome,expression,and ginsenoside content databases,this study employed bioinformatic methods to systematically analyze the genes encoding SQE in ginseng.We first selected six PgSQE candidates that were closely involved in ginsenoside biosynthesis and then identified PgSQE08-01 to be highly associated with ginsenoside biosynthesis.Next,we constructed the overexpression vector pCAMBIA3301-PgSQE08-01 and the RNAi vector pART27-PgSQE08-01 and transformed ginseng adventitious roots using Agrobacterium rhizogenes,to obtain positive hairy-root clones.Thereafter,quantitative reverse transcriptionpolymerase chain reaction and high-performance liquid chromatography were used to determine the expression of relevant genes and ginsenoside content,respectively.Then,we focused on the function of PgSQE08-01 gene,which was noted to be involved in ginsenoside biosynthesis.Thus,these findings not only provided a molecular basis for the identification of important functional genes in ginseng but also enriched genetic resources for the biosynthesis of ginsenosides using synthetic biology.

High-throughput transcriptional profiling of perturbations by Panax ginseng saponins and Panax notoginseng saponins using TCM-seq

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.

Comparative analysis of physicochemical properties,ginsenosides content andα-amylase inhibitory effects in white ginseng and red ginsen

Ginseng(Panax ginseng C.A.Meyer)as a common dietary adjunct is widely applied in Traditional Chinese Medicine due to its health-promoting properties,but the differences between white ginseng and red ginseng was rarely studied.In the present study,color parameters and scanning electron microscope(SEM)were determined to evaluate the differences of ginseng color and microstructure induced by processing procedure.Quantitative analysis of multi-components by a single-marker(QAMS)method and anti-α-amylase activity test were used to assess variations of chemical ingredients and pharmacological activity between white and red ginseng.Finally,molecular docking studies were carried out to screen out the most effective compound againstα-amylase.Results indicated that processing had a significant impact on the physicochemical properties and pharmacological activity of white and red ginseng.After processing,the color value of L*declined significantly.Red ginseng sample displayed a compact structure and presented of a gel layer on the surface compared to white ginseng.Additionally,the content of ginsenosides and the activity of anti-α-amylase decreased.The contents of total ginsenosides were positively correlated with the anti-α-amylase activities of ginseng,and ginsenoside Rb1 might be the most effective compound to inhibit the activity ofα-amylase.

Molecular Mechanism of Ginseng in Treating Nephrotic Syndrome Based on Network Pharmacology and Experimental Verification

[Objectives]To study the potential molecular mechanism of ginseng in treating nephrotic syndrome(NS)by using network pharmacology,molecular docking and experimental verification methods.[Methods]The active components and targets of ginseng were obtained through the network pharmacology database,and the potential targets for the treatment of NS were predicted.The STRING data platform and Cytoscape software were used to construct protein interaction network,and carry out GO and KEGG enrichment analysis.Molecular docking of active components of ginseng and core targets was performed.The in vitro experiment verified the improvement effect of kaempferol,a key active ingredient of ginseng,on podocyte injury.[Results]After screening,17 active components of ginseng and 38 key targets for treating NS were obtained.GO and KEGG enrichment analysis showed that NF-κB,MAPK and other inflammatory pathways were involved.Molecular docking results show that the core components had good binding activity to key targets.The results of in vitro experiments show that kaempferol can reduce the phosphorylation level of AKT1,down-regulate the expression levels of NF-κB p65 and p-NF-κB p65,play an anti-inflammatory effect by inhibiting the activation of NF-κB pathway,and improve podocyte injury.[Conclusions]Ginseng may play a role in the treatment of NS by regulating multiple targets and pathways such as inflammatory response,substance metabolism,and signal transduction.

Effects of Ginseng Protein on Gut Microbiota and BDNF/TrkB Signaling Pathway in Alzheimer s Disease Mice

[Objectives] To investigate the effects of ginseng protein on gut microbiota and BDNF/TrkB signaling pathway in Alzheimer s disease (AD) mice. [Methods] D-galactose/AlCl 3 co-induction was used to establish AD model, and mice were randomly divided into normal group 1, normal group 2, model group 1, model group 2, ginseng protein group, and microbiota transplantation group. Morris water maze experiment was used to evaluate learning and memory ability, and Western blot method was used to detect the expression of APP, p-Tau, BDNF, TrkB, p-TrkB proteins in brain tissue, and 16S rDNA was used to detect diversity of fecal microbiota. [Results] Ginseng protein and microbiota transplantation can shorten the escape latency of mice ( P <0.05), increase the number of crossing platforms ( P <0.05), reduce the expression of APP and p-Tau proteins in brain tissue ( P <0.05, P <0.01), increase the expression of BDNF, p-TrkB, p-TrkB/TrkB proteins ( P <0.05, P <0.01), and reduce the abundance of Alloprevotella, Ruminococcaceae _UCG-014, Prevotellaceae _UCG-001, and Ruminococcus _1 ( P <0.05, P <0.01). [Conclusions] The action mechanism of ginseng protein anti AD may be through regulating gut microbiota diversity and activating the BDNF/TrkB signaling pathway.

Effects of ginseng powder on chemical composition, antioxidant and sensory properties of biscuits

This work aimed to evaluate the quality of biscuits incorporated with ginseng powder.This powder was incorporated into wheat flour at levels ranging from 0 to 4%and used in the production of biscuits.The biscuits were analysed for proximate composition,sensory attributes,phytochemical content,and antioxidant properties.Results revealed that the moisture and fat contents of the biscuits significantly decreased with the incorporation of ginseng powder.Conversely,protein(3.15 to 4.55 g/100 g DM),carbohydrate(27.80 to 31.10 g/100 g DM)and fiber contents(6.21 to 13.74 g/100 g DM)significantly increased with incorporation.The total polyphenols,saponins,and total flavonoid contents increased from 1.67 to 1.99 mgGAE/100 g DM,0 to 0.69 mg,and 0.87 to 1.83 QE/100g DM respectively.Ginseng powder also significantly increased the DPPH scavenging activity,from 42.05%to 52.62%,and FRAP value from 2.25 to 7.71μgGAE/g with increased incorporation.However,the sensory attributes of the biscuits were not affected by the incorporation of ginseng.Ginseng powder can therefore be incorporated in wheat flour at up to 4%level for biscuits production as a functional food for the promotion of human health.

Antagonistic Effects of Sphingomonas and Pseudomonas aeruginosa on 4 Kinds of Pathogenic Bacteria of Ginseng

[Objectives]To explore effective biocontrol methods for diseases in the process of ginseng cultivation,and develop an efficient and environmentally friendly biocontrol agent.[Methods]In this study,2 strains were isolated from biogas slurry,and Cylindrocarpon destructans(XF),Fusarium solani(GF),Botrytis cinerea Pers(HM)and Alternaria panax Whetz(HB)were used as test materials.The strains were isolated and identified by dilution plate method,16S rDNA sequence identification method,confrontation culture method,filter paper method and ultraviolet spectrophotometer method,and the bacteriostatic activity and bacteriostatic rate were tested.[Results]Strain 15(Sphingomonas)and strain 19(Pseudomonas aeruginosa)were screened out through identification and analysis,and they grew stably within 8-10 d.The bacteriostatic rates of strain 15 against A.panax and B.cinerea were 47.37%and 43.40%,respectively,and the bacteriostatic rates of strain 19 against A.panax and B.cinerea were 62.30%and 63.27%,respectively.The bacteriostatic activity of the extract of strain 19 increased with the increase of OD_(600) value,and the bacteriostatic effect was optimal when the OD_(600) value was in the range of 0.8-1.0,up to 70%,so it had a strong biocontrol potential.[Conclusions]This experiment provides convenience for more effective inoculation,establishes a fast,simple and accurate method for the determination of the best bacteriostatic rate of P.aeruginosa culture solution to HM,and lays a foundation for large-scale culture of P.aeruginosa culture solution.Besides,it is expected to provide a theoretical basis for the efficient control of ginseng B.cinerea in field production,use it for the prevention and control of ginseng shoot diseases,and provide a reference for the efficient and diverse development of biocontrol agents for ginseng shoot diseases.

Immunoregulation Effect of Red Ginseng Aqueous Extract on Mice

[Objectives]The paper was to observe the effects of red ginseng aqueous extract on immune function of mice.[Methods]The aqueous extract of red ginseng was obtained by water extraction and concentration,and 10 mL of aqueous extract was equivalent to 2.86 g of raw materials.Total 240 mice were randomly divided into blank control group,low dose,medium dose and high dose groups of red ginseng aqueous extract,60 mice each group.Mice in low dose,medium dose and high dose groups were intragastrically administered with 0.24,0.48,1.43 g/kg red ginseng aqueous extract once a day respectively,and those in blank control group were intragastrically administered with equal volume of deionized water at the dose of 0.1 mL/10 g once a day for consecutive 30 d.The immunoregulation effects of red ginseng aqueous extract on mice were explored by organ/body weight ratio measurement,delayed type hypersensitivity(DTH)reaction,Con A-induced spleen T lymphocyte transformation test,antibody-producing cells test,half hemolytic value(HC 50)test,carbon particle clearance test,phagocytosis test of chicken red blood cells by macrophages and NK cell activity test.[Results]Compared with the blank control group,the degree of toe swelling in low dose group and medium dose group of red ginseng aqueous extract[(0.62±0.14)mm,(0.53±0.12)mm vs.(0.36±0.10)mm]significantly increased(P<0.05).The ability of T lymphocytes proliferation(0.173±0.054,0.189±0.063 vs.0.098±0.012)in low dose group and high dose group significantly increased(P<0.05).The number of haemolytic plaque(137.49×10^(3)±24.73×10^(3),148.43×10^(3)±27.53×10^(3) vs.112.96×10^(3)±26.28×10^(3))in medium dose group and high dose group significantly increased(P<0.05).The phagocytosis rate(35.67%±3.82%,49.26%±6.54%,57.92%±7.36%vs.24.34%±4.22%)and phagocytosis index(0.72±0.23,0.82±0.15,0.91±0.26 vs.0.35±0.11)of low dose,medium dose,high dose groups significantly increased(P<0.05).However,there was no statistical difference in spleen/body weight ratio,thymus/body weight ratio,half hemolytic value,carbon particle clearance and NK cell activity.[Conclusions]Red ginseng aqueous extract could enhance the immunity of mice.

Community Structure of Endophytic Fungi in Ginseng and Its Correlation Analysis with Active Components

[Objectives]The paper was to explore the correlation between active components of ginseng and community structure of endophytic fungi in ginseng under different cultivation methods.[Methods]Using ginsengs under two main planting patterns in Huanren County,Liaoning Province as the samples,the community structure of endophytic fungi was studied by molecular means,and the chemical components were identified and the content of chemical markers was measured via LC-MS.Moreover,the correlation between medicinal components of ginseng and diversity of endophytic fungal flora was analyzed.[Results]The diversity and active components of endophytic fungi of ginseng were different.The diversity analysis showed that the medicinal components of ginseng were positively correlated with some fungal groups in ginseng.[Conclusions]Artificial intervention in the planting process would affect the endophytic fungi in medicinal materials,and further affect the chemical components of ginseng medicinal materials.

A Survey of the History and Applications of American Ginseng(西洋参)

American ginseng(Panax Quinquefolium L),also known as Xi Yang Shen(西洋参 Radix Panacis Quinquefolii),is indigenous to the United States and Canada.Its discovery in the 1700s paved the way for international trade,which boosted the economy of the New World and helped build commercial ties between the US and China.Due to its therapeutic effects,the demand for American ginseng grew steadily,eventually causing the volume of wild ginseng to dwindle.As a result,laws and regulations were introduced requiring farmers and exporters to engage in harvesting practices that would preserve wild American ginseng.Nowadays,wild American ginseng is considered an endangered and protected species.This article discusses the history,properties,and applications of American ginseng to optimize its use and protection.

人参(Panax ginseng)根系分泌物成分对人参致病菌的化感效应

采用室内培养结合生物学测定的试验方法,研究了不同浓度人参(Panax ginseng)根系分泌物成分苯甲酸、邻苯二甲酸二异丁酯、十六酸和2,2二-(4羟-苯基)丙烷对人参立枯丝核菌(Rhizoctonia solani)、黑斑菌(Alternaria panax)、疫病菌(Phytophthora cactorum)、菌核菌(Sclerotinia schinseng)、锈腐菌(Cylindrocarpon destructans)和绿色木霉菌(Trichoderma viride)菌落生长及孢子萌发的化感效应。结果显示,不同浓度人参根系分泌物成分对人参致病菌及绿色木霉菌的化感效应存在显著差异。苯甲酸浓度与人参立枯丝核菌、菌核菌和锈腐菌菌落生长以及人参黑斑菌、锈腐菌孢子萌发呈负相关,与人参黑斑菌、绿色木霉菌菌落生长呈正相关;对人参疫病菌菌落生长的化感效应表现为低浓度和高浓度抑制,中浓度促进。邻苯二甲酸二异丁酯浓度与人参立枯丝核菌、黑斑菌、菌核菌和绿色木霉菌菌落生长以及人参黑斑菌孢子萌发呈负相关;对人参锈腐菌菌落生长和孢子萌发表现为低浓度和高浓度抑制,中浓度促进;对人参疫病菌菌落生长表现为低浓度和中浓度抑制,高浓度促进。2,2二-(4羟-苯基)丙烷浓度与人参立枯丝核菌、黑斑菌、疫病菌、绿色木霉菌菌落生长以及人参黑斑菌、锈腐菌孢子萌发呈负相关;对人参菌核菌、锈腐菌菌落生长表现中浓度促进,高浓度抑制。十六酸浓度与人参锈腐菌、疫病菌和绿色木霉菌菌落生长呈正相关,与人参锈腐菌孢子萌发呈负相关,对黑斑菌孢子萌发表现为中浓度抑制。4种根系分泌物的等量混合物浓度与人参致病菌及拮抗木霉菌菌落生长速率呈负相关。

Research on the Extraction Techniques of Ginseng Based on Patent Analysis

Objective To analyze the gaps between patents of ginseng extraction method in China and South Korea,and to provide research and development directions for Chinese companies.Methods By observing the changes in the amount of patent applications over time,the distribution of application countries and regions,the analysis of applicants,the extraction of technical topics,and the development and change of method,an in-depth discussion on the patent of ginseng extraction method in China and South Korea was conducted.Results and Conclusion China lacks the mastery of advanced technology and the exploration of popular fields,while South Korea is just the opposite of China.Therefore,Chinese enterprises should focus on extraction processes and actively explore new technical fields.

人参(Panax ginseng)的氮过剩现象──人参氮毒原因初探

1985-1990年间，我们在长白县及公主岭进行的四次NPK三要素试验中，发现氮肥抑制了人参生长发育，降低了产量（5％显著水准），而倍量氮肥，减产达1％显著水平。继续研究证明，人参硝酸还原酶活力（NRA）甚低，追施氮肥使组织，积累，促进了呼吸作用，植株干物中N、Si及Cu、Zn、Fe、Mn含量增加，Ca、Mg、K、Na等阳离子减少，表现出氮毒征象。本文称氮过剩症，它是指氮素供应超过了由极低NRA催化的转化代谢速率，而引起的一种毒害现象。

Ginsenoside-Rg_6, a Novel Triterpenoid Saponin from the Stem-Leaves of Panax ginseng C. A. Mey.

A novel dammarane-type triterpene oligoglycoside, named ginsenoside-Rg6 3, was isolated from the stem-leaves of Panax ginseng C. A. Mey., together with two known ones, 20(S)-ginsenoside-Rg2 1 and 20(R)-ginsenoside-Rg2 2. On the basis of chemical and physicochemical evidence , the structure of ginsenoside-Rg6 have been elucidated as 6-O-(-L-rhamnosyl-(1?2)-(-D-glucopyranosyl-dammarane-(E)-20(22), 24-diene-3(, 6(, 12(-triol.

Korean red ginseng promotes hippocampal neurogenesis in mice

Neurogenesis in the adult hippocampus plays a major role in cognitive ability of animals including learning and memory.Korean red ginseng (KRG) has long been known as a medicinal herb with the potential to improve learning and memory;however,the mechanisms are still elusive.Therefore,we evaluated whether KRG can promote cognitive function and enhance neurogenesis in the hippocampus.Eight-week-old male C57BL/6 mice received 50 mg/kg of 5-bromo-2′-deoxyuridine (BrdU) intraperitoneally and 100 mg/kg of KRG or vehicle orally once a day for 14 days.Pole,Rotarod and Morris water maze tests were performed and the brains were collected after the last behavioral test.Changes in the numbers of BrdU- and BrdU/ doublecortin (DCX;a marker for neuronal precursor cells and immature neurons)-positive cells in the dentate gyrus and the gene expression of proliferating cell nuclear antigen (a marker for cell differentiation),cerebral dopamine neurotrophic factor and ciliary neurotrophic factor in the hippocampus were then investigated.KRG-treated mice came down the pole significantly faster and stood on the rotarod longer than vehicle-treated mice.The Morris water maze test showed that KRG administration enhanced the learning and memory abilities significantly.KRG also significantly increased BrdU- and BrdU/DCX-positive cells in the dentate gyrus as well as the proliferating cell nuclear antigen,cerebral dopamine neurotrophic factor and ciliary neurotrophic factor mRNA expression levels in the hippocampus compared to vehicle.Administration of KRG promotes learning and memory abilities,possibly by enhancing hippocampal neurogenesis.This study was approved by the Pusan National University Institutional Animal Care and Use Committee (approval No.PNU-2016-1071) on January 19,2016.

Study on Agricultural Intellectual Property Protection and High-Quality Development of Traditional Chinese Medicine of Ginseng

Agricultural intellectual property has the functions of carrying forward agricultural cultural heritage,encouraging agricultural scientific and technological innovation,cultivating the brand of agricultural products and so on. The protection and application of agricultural intellectual property rights can enhance the vitality,innovation and competitiveness of agricultural economy,and promote the strategy of high-quality agricultural development and rural revitalization. Ginseng is a traditional medicinal variety with superior characteristics in Northeast China,and it is one of the most famous " three treasures" in Northeast China. This paper introduces ginseng( Panax ginseng C. A. Mey.) and its medicinal value,analyzes the traditional knowledge,biological genetic resources,new plant varieties,patents,geographical indications,trademarks and other agricultural intellectual property resources of ginseng,studies the traditional Chinese medicine GAP certification and certification of " Three Agricultural Products" and the construction of the dominant area of agricultural product( Fusong ginseng) with Chinese characteristics,and explores the rural revitalization strategy and traditional Chinese medicine development strategy,ginseng agricultural intellectual property protection and high quality development strategy.

Comparing Ginsenoside Production in Leaves and Roots of Wild American Ginseng (<i>Panax quinquefolius</i>)

American ginseng, Panax quinquefolius L., is an herbaceous perennial species that is destructively harvested for its bioactive compounds called ginsenosides. The demand for this herb fosters illegal poaching and over-harvesting that reduces genetic variability and population viability. Five wild populations in western North Carolina were studied to better understand the production of ginsenosides in leaf and root tissues. Total ginsenoside concentration was significantly higher in leaves than roots, though total yield was higher in roots due to greater root biomass. However, some ginsensosides (Rb2, Rd and Re) had higher or more consistent yields in leaves than roots, so might be developed into a sustainable source of these medicinally-active compounds. Additionally, we identified regional root chemotypes that differed in the production of the ginsenosides Rg1 and Re and could be developed into regional cultivars depending on the desired panel of ginsenosides.

Inhibitory Effects of Ginseng Water Extract on Hydroxyl Radical

[Objectives] The aim of this work was to compare the inhibitory effects of different water-soluble extracts of ginseng on hydroxyl radical. [Methods]The water-soluble effective parts of ginseng were extracted by orthogonal test,the hydroxyl radical scavenging rate of each extract was determined by salicylic acid method,and the extraction process of the optimum hydroxyl radical inhibitor was optimized using variance analysis and cluster analysis. [Results] A total of 16 samples were prepared. Among them,the sample No. 12 had the highest hydroxyl radical scavenging rate,and the inhibition rate reached 105. 08% at the concentration of 1 g/L. [Conclusions] Ginseng water extract contained hydroxyl radical scavenger,of which the optimum extraction process was as follows: p H of 8,solid to liquid ratio of 1∶ 10,extraction temperature of 4℃,extraction time of 12 h and extraction times of 3.

Use of Network Pharmacology and Molecular Docking to Investigate the Mechanism by Which Ginseng Ameliorates Hypoxia

Hypoxia is a common pathological process in various clinical diseases and is characterized by abnormal changes in metabolism,function,and morphological structure of tissues resulting from insufficient oxygen supply or oxygen barriers in tissues.In particular,hypoxia in vital organs such as the brain and heart is an important cause of death[1].The prevention of tissue hypoxia and the treatment of hypoxia-induced tissue damage are urgent issues.

Microbial community diversity analysis of <i>Panax ginseng</i>rhizosphere and non-rhizosphere soil using randomly amplified polymorphic DNA method

DNA sequence diversities of soil microbial communities in rhizosphere and non-rhizosphere of 1 - 6 years Panax ginseng were evaluated by random amplified polymorphic DNA (RAPD). Total genomic DNA of soil samples were amplified by twenty-four out of two hundred random primers. The products were separated in agarose gel, and 359 and 181 reliable fragments were generated, of which, 324 and 123 were polymorphic. The gene diversity index, Shannon’s information index, and similarity coefficient were calculated, and results showed that, diversity of soil microbial community at DNA level was present. Furthermore, with the increasing of P. ginseng growing at one site, deference on soil microbial community DNA sequence between rhizosphere and non-rhizosphere tend to be more and more significant. In addition, RAPD technique was proved to be an effective tool to assess the diversity of soil microbial communities. From results we deduced that, the ecological function that performed by soil microbes may changed following the change of microbial community in rhizospheric and non-rhizospheric soils, and finally broke the balance of soil nutrition and energy cycle, soil spatial structure, microbial ecology etc.