High-throughput transcriptional profiling of perturbations by Panax ginseng saponins and Panax notoginseng saponins using TCM-seq / 药物分析学报

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 sa-ponins(PNS)and Panax ginseng saponins(PGS)extracted from PN and PG,respectively.We also char-acterized 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 tran-scriptional 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.

Ecological Suitability and Quality Zoning of Panax ginseng Cultivated Under Forest in Liaoning Province / 中国实验方剂学杂志

ObjectiveTo identify potential distribution areas for wild Panax ginseng cultivated under forest in Liaoning province， China， and analyze the ecological factors of spatial stratified heterogeneity affecting its ecological suitability and quality suitability. MethodWild Panax ginseng samples cultivated under forest were collected from 33 cultivation bases in Liaoning province. The Maxent maximum entropy model and ArcGIS were used to delineate the ecological suitability zones. Correlation analysis was performed on seven indicators and 110 ecological factors. Variables with significant correlation （P<0.05） were used to build partial least squares regression analysis models. A comprehensive quality zoning was conducted using the analytic hierarchy process （AHP）. The geographic detector was employed to analyze the interactions among dominant ecological factors of spatial stratified heterogeneity affecting habitat suitability， quality suitability， and the ecological driving factors. ResultVegetation type was the most influential ecological factor for delineating the ecological suitability zones for wild Panax ginseng in Liaoning province. The main ecological suitability areas for wild Panax ginseng cultivated under forest were located in the northeast， east， and southeast regions along the line from Xifeng County to Gaizhou City. The comprehensive quality suitability of wild Panax ginseng cultivated under forest was highest in Kuandian County and Huanren County and gradually decreased to the northwest and southwest. Within the delineated regions， the suitability conditions and comprehensive quality of wild Panax ginseng cultivated under forest were primarily influenced by the interactions between radiation and precipitation factors. The content of the measured samples was significantly higher than the standards in the 2020 edition of the Chinese Pharmacopoeia， indicating the high overall quality of wild Panax ginseng in Liaoning Province. ConclusionAccording to the zoning and prediction results， areas in Fengcheng City， Xiuyan County， Zhuanghe City， Liaoyang County， Tieling County， Xifeng County， Gaizhou City， Haicheng City， and Dashiqiao City showed large potential distribution areas with high quality， making them highly promising for wild Panax ginseng cultivation. However， further experimental verification is required. The zoning results can provide insights for research on habitat suitability and comprehensive quality accumulation of wild Panax ginseng cultivated under forest， as well as guidance for the search for potential cultivation areas and industrial development of wild Panax ginseng in Liaoning Province.

Effects of total ginsenosides from Panax ginseng stems and leaves on gut microbiota and short-chain fatty acids metabolism in acute lung injury mice / 中国中药杂志

This study aimed to investigate the biological effects and underlying mechanisms of the total ginsenosides from Panax ginseng stems and leaves on lipopolysaccharide(LPS)-induced acute lung injury(ALI) in mice. Sixty male C57BL/6J mice were randomly divided into a control group, a model group, the total ginsenosides from P. ginseng stems and leaves normal administration group(61.65 mg·kg~(-1)), and low-, medium-, and high-dose total ginsenosides from P. ginseng stems and leaves groups(15.412 5, 30.825, and 61.65 mg·kg~(-1)). Mice were administered for seven continuous days before modeling. Twenty-four hours after modeling, mice were sacrificed to obtain lung tissues and calculate lung wet/dry ratio. The number of inflammatory cells in bronchoalveolar lavage fluid(BALF) was detected. The levels of interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in BALF were detected. The mRNA expression levels of IL-1β, IL-6, and TNF-α, and the levels of myeloperoxidase(MPO), glutathione peroxidase(GSH-Px), superoxide dismutase(SOD), and malondialdehyde(MDA) in lung tissues were determined. Hematoxylin-eosin(HE) staining was used to observe the pathological changes in lung tissues. The gut microbiota was detected by 16S rRNA sequencing, and gas chromatography-mass spectrometry(GC-MS) was applied to detect the content of short-chain fatty acids(SCFAs) in se-rum. The results showed that the total ginsenosides from P. ginseng stems and leaves could reduce lung index, lung wet/dry ratio, and lung damage in LPS-induced ALI mice, decrease the number of inflammatory cells and levels of inflammatory factors in BALF, inhibit the mRNA expression levels of inflammatory factors and levels of MPO and MDA in lung tissues, and potentiate the activity of GSH-Px and SOD in lung tissues. Furthermore, they could also reverse the gut microbiota disorder, restore the diversity of gut microbiota, increase the relative abundance of Lachnospiraceae and Muribaculaceae, decrease the relative abundance of Prevotellaceae, and enhance the content of SCFAs(acetic acid, propionic acid, and butyric acid) in serum. This study suggested that the total ginsenosides from P. ginseng stems and leaves could improve lung edema, inflammatory response, and oxidative stress in ALI mice by regulating gut microbiota and SCFAs metabolism.

Sensitivity baseline establishment and resistance risk assessment of Botrytis cinerea from Panax ginseng to prochloraz / 中国中药杂志

This study aimed to establish the baseline sensitivity of Botrytis cinerea from Panax ginseng to prochloraz, and ensure the fitness of prochloraz-resistant mutants and the cross-resistance of B. cinerea to prochloraz and commonly used fungicides for the prevention and control of gray mold including boscalid, pyraclostrobin, iprodione, and pyrimethanil. The sensitivity of B. cinerea from P. ginseng to fungicides was determined by the mycelial growth rate method. The prochloraz-resistant mutants were screened out through fungicide domestication and ultraviolet(UV) induction. The fitness of resistant mutants was determined through the stability of subculture, mycelial growth rate, and pathogenicity test. The cross-resistance between prochloraz and the four fungicides was determined by Person correlation analysis. The results showed that all B. cinerea strains tested were sensitive to prochloraz, and the EC_(50) value ranged from 0.004 8 to 0.062 9 μg·mL~(-1), with an average of 0.022 μg·mL~(-1). The sensitivity frequency distribution diagram showed that 89 B. cinerea strains were located within the main peak with a continuous single peak curve, and the average EC_(50) value of 0.018 μg·mL~(-1) was taken as the baseline sensitivity of B. cinerea to prochloraz. The fungicide domestication and UV induction obtained 6 resistant mutants, among which 2 strains were unstable and the other 2 strains showed decreased resistance after multiple generations of culture. Furthermore, the mycelial growth rate and spore yield of all resistant mutants were lower than those of their parents, and the pathogenicity of most mutants was lower than that of their parents. In addition, prochloraz had no obvious cross-resistance with boscalid, pyraclostrobin, iprodione, and pyrimethanil. In conclusion, prochloraz has great potential for controlling gray mold in P. ginseng, and the resistance risk of B. cinerea to prochloraz is low.

Transcriptome analysis and differential gene screening of Panax ginseng seed in different sand storage stages / 药学学报

Analyze the changes in gene expression profiles during the process of Panax ginseng seed dormancy release, and screen for differential genes, providing a basis for analyzing the mechanism of P. ginseng seed dormancy release. Comparative transcriptome analysis was conducted by using RNA-Seq sequencing technology in P. ginseng seeds stored at different low temperature. A total of 80.97 Gb of Raw reads and 80.19 Gb of Clean reads were obtained from the transcriptome. Principal component analysis and correlation analysis showed that there were significant differences in gene expression patterns at different developmental stages. Upset results showed that 46 248 unigenes were co-expressed in four stages, and 414, 445, 400 and 389 unigenes were specifically expressed in 0, 8,14 and 28 days, respectively. Gene Ontology functional annotation showed that the differentially expressed genes were mainly involved in nsaturated fatty acid biosynthetic process, nuclear body and oxidoreductase activity. Encyclopedia of Genes and Genomes metabolic pathway showed that differentially expressed genes were mainly involved in peroxisome, mitogen-activated protein kinase signaling pathway-plant, plant hormone signal transduction, ribosome, biosynthesis of unsaturated fatty acid, circadian rhythm-plant and other metabolic pathways. In the process of P. ginseng seed dormancy release, multiple biological processes, such as unsaturated fatty acid biosynthesis and plant hormone signal transduction, are required to coordinate regulation, which constitutes a complex dormancy release regulation network. Transcriptome analysis and differential gene screening of P. ginseng seeds at different sand storage time laid a foundation for the analysis of P. ginseng seed dormancy release mechanism and molecular breeding.

Comparative analysisof Saponins in different medicinal parts of Panax Ginseng / 国际中医中药杂志

Objective:To analyze the differences of Saponins in different parts of Panax ginseng, which couldprovide reference for a comprehensive quality evaluation.Methods:UFLC- Triple-TOF-MS/MS was used to analyze the Saponins in Ginseng Radix et Rhizoma, Ginseng Folium and Ginseng Flos,The analysis was carried out on a SynergiTM Hydro-RP 100A column, Gradient elution of water (containing 0.1% formic acid) (A)-acetonitrile (B). Principal component analysis (PCA) was used to analyze the grouping of samples, and partial least squares regression (PLS-DA) was used to classify the samples to find the differences of chemical components in different medicinal parts of Panax ginseng. Significant differences in saponins and its rules were found by multivariate statistical analysis.Results:PCA indicated that there was remarkable difference in saponins of Ginseng Radix et Rhizoma, Ginseng Folium and Ginseng Flos, ten different components were found by PLS-DA. Conclusion:There exists obvious differences of different medicinal parts of Panax Ginseng which could provide foundation for the further research and rathional use of Panax ginseng.

Metabolomics analysis reveals the renal protective effect of Panax ginseng C. A. Mey in type 1 diabetic rats / 中国天然药物

The dry root and rhizome of Panax ginseng C. A. Mey has garnered much interest owing to its medicinal properties against diabetes and cardiovascular diseases. In this study, an ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS)-based metabolomics approach was used to illustrate the therapeutic mechanisms of ginseng extract on the serum and urinary metabolic profiles in streptozotocin-induced type 1 diabetes mellitus (T1DM) rats. Pharmacological and renal parameters in response to the administration of ginseng were also evaluated. In total, 16 serum endogenous metabolites and 14 urine endogenous metabolites, including pyruvic acid, indoleacetic acid, and phenylacetylglycine, were identified as potential biomarkers for diabetes. Pathway enrichment and network analysis revealed that the biomarkers modulated by ginseng were primarily involved in phenylalanine and pyruvate metabolism, as well as in arginine biosynthesis. Moreover, the levels of several renal injury-related biomarkers in T1DM rats were significantly restored following treatment with ginseng. The administration of the extract helped maintain tissue structure integrity and ameliorated renal injury. The findings suggest that the regulatory effect of ginseng extract on T1DM involves metabolic management of diabetic rats, which subsequently attenuates T1DM-induced early renal dysfunction.

Establishment of content determination of 5 saponins in Huoxue zhitong capsules/tablets and confirmation of illegal addition / 中国药房

OBJECTIVE To establish a method for simultaneous determination of 5 saponins in Huoxue zhitong capsules/ tablets and to confirm the illegal addition of Panax ginseng ，Panax quiquefolium and stems and leaves of Panax notoginseng . METHODS Ultra-high performance liquid chromatography tandem mass spectrometry was used . The Agilent RRHD Eclipse Plus C18 column was used with mobile phase of water （containing 0.1% formic acid ）-acetonitrile for gradient elution at a flow rate of 0.35 mL/min. The column temperature was 35 ℃andthesamplesizewas 2 μL. Using electrospray ionization source ，negative ion scanning was carried out in multi -reaction monitoring mode . RESULTS The linear ranges of notoginsenoside R 1，ginsenoside Rb 1， ginsenoside Rg 1，ginsenoside Rd ，ginsenoside Re ，ginsenoside Rf （an unique ingredient of P. ginseng），ginsenoside Rb （3 an unique ingredient of stems and leaves of P. notoginseng）and pseudo -ginsenoside F 11（an unique ingredient of P. quiquefolium）were 9.99- 1 499.50，9.99-1 499.50，10.01-1 500.80，9.99-1 499.10，10.00-1 500.20，9.99-1 499.50，10.01-1 500.80，9.99-1 499.00 ng/mL （R2＞0.997）；the detection limits and the quantitative limits were not higher than 2.64 and 8.06 ng/mL，respectively. RSDs of precision，repeatability and stability （24 h）tests were all less than 6%. The average recoveries of saponins in capsules and tablets were 98.72%-102.40% and 95.18%-106.47%，respectively（all RSDs ＜5%，n＝6）. In 18 batches of Huoxue zhitong capsules ，the contents of ginsenoside Re ，ginsenoside Rd ，ginsenoside Rg 1，notoginsenoside R 1 and ginsenoside Rb 1 were 291.79-426.89，427.71- 677.49，2 294.28-3 371.43，571.22-848.19 and 1 841.33-2 959.12 μg/g，respectively；the contents of ginsenoside Rb 3 were no more than 45.02 μg/g. In 22 batches of Huoxue zhitong tablets，the contents of above indicators of P. notoginseng were 44.11-393.83，80.48-549.55，393.36-3 548.57，79.83- 872.60，and 288.64-2 912.66 μg/g，respectively；the contents of ginsenoside Rb 3 were no more than 44.79 μg/g. Ginsenoside Rf and pseudo -ginsenoside F 11 were not detected in the two preparations. CONCLUSIONS The method can be used to determine the contents of saponins in Huoxue zhitong preparations . No illegal addition of P. ginseng and P. quiquefolium are found in 40 batches of preparations ，but the input of P. notoginseng in some batches of tablet samples is less .

Transcriptome-wide identification and expression pattern analysis for Dof gene family in Panax ginseng from Jilin / 中国中药杂志

Dof(DNA binding with one finger), a unique class of transcription factors in plants, play an important role in seed development, tissue differentiation, and metabolic regulation. To identify the number and function of Dof gene family members in Panax ginseng, this study identified the members of Dof gene family in P. ginseng and systematically analyzed their structures, evolution, functional differentiation, expression patterns, and interactions using bioinformatics methods at the transcriptome level. At the same time, the association analysis of Dof genes from P. ginseng with key enzyme genes for ginsenoside synthesis was carried out to screen the candidate PgDof genes involved in the regulation of ginsenoside biosynthesis. The results showed that there were 54 genes belonging to the Dof gene family in P. ginseng from Jilin. All PgDof genes had Zf-Dof conserved motifs, implying that they were evolutionarily conserved and could be divided into five groups. Expression pattern analysis confirmed that the expression of PgDof gene family members in different tissues, different year-old P. ginseng, and different farm varieties varied significantly. Simultaneously, as revealed by "gene-saponin content" and "gene-gene" linkage analysis, an important candidate PgDof14-1 gene involved in the regulation of ginsenoside biosynthesis was obtained. From the established genetic transformation system of this gene in the hairy roots of P. ginseng, a positive hairy root clone was determined. This study has laid a theoretical foundation for the study of Dof gene family in P. ginseng.

Optimal Potassium Application Rate for Panax ginseng / 中国实验方剂学杂志

ObjectiveThe law of fertilizer requirement serves as the basis for the fertilization of medicinal plants， development of special fertilizer， and high-quality medicinal materials. MethodThis study aims to explore the optimal potassium application rate for Panax ginseng to achieve high yield and quality of the medicinal material and targeted management of potassium fertilizer. To be specific， 6 concentration gradients （0， 2， 4， 8， 10， and 12 mmol·L-1） of potassium sulfate （potassium fertilizer） were designed and applied to the 4-year-old P. ginseng in CK， C1， C2， C3， C4， and C5 treatments， respectively. Thereby， the influence of potassium concentration on P. ginseng was observed. ResultWhen potassium sulfate was applied at 8 mmol·L-1， P. ginseng had the chlorophyll content of 32.13%， net photosynthetic rate of 2.548 8 µmol·m-2·s-1， and activities of superoxide dismutase （SOD）， peroxidase （POD）， and catalase （CAT） significantly higher than those in the CK， C1， C4， and C5 treatments （P<0.05）. The average fresh weight of P. ginseng roots was 6.25 g， 134% up from the CK， and the content of ginsenoside Rg1 （5.24 mg·g-1） and Re （4.17 mg·g-1） and total saponins （12.33 mg·g-1） was significantly higher than that in CK and other treatments （P<0.05）. Thus， 8 mmol·L-1 potassium sulfate was most favorable for the growth and effective component accumulation of four-year-old P. ginseng. ConclusionThis study expounds the effect of potassium fertilizer on the yield and quality of P. ginseng， which is expected to help guide the precise application of potassium fertilizer in P. ginseng production in the field and lay a theoretical basis for the development of special fertilizer for P. ginseng and the optimization of fertilization technology.

Yield， Quality， and Soil Microecology of Panax ginseng Under Different Forests / 中国实验方剂学杂志

ObjectiveTo explore the forest type and soil environment suitable for Panax ginseng. MethodThe yield， quality， soil chemical properties， soil enzyme activity， and soil microbial metabolism of 9-year-old P. ginseng under different forests were investigated. ResultThe quality of P. ginseng was significantly different among forest types. To be specific， P. ginseng under the Quercus mongolica forest had the highest quality， with the total saponin content of 2.27% which was 51.89% higher than that in P. ginseng under Larix gmelinii forest. The yield of P. ginseng under Q. mongolica forest and L. gmelinii forest （30 g·m-2） was the highest， 62.5% higher than that under Betula platyphylla forest. The soil content of organic matter， Cu， and Zn， and activity of sucrase and urease under Q. mongolica forest were lower than those under other forest types. The utilization rate of D-galacturonic acid by soil microorganisms under Q. mongolica forest was higher than that under other forest types， but the utilization rate of L-phenylalanine was lower than that under other forest types. The utilization rate of 2-hydroxybenzoic acid by soil microorganisms of B. platyphylla forest was significantly lower than that under other forest types. There was a negative correlation between soil Zn and ginsenoside Rb1 and Rc， and between soil K and ginsenoside Rb2 and Rb3. Mn and Cu were positively correlated with most saponins. The results of redundancy analysis showed that the soil microorganisms using carbon sources of amino acids， esters， acids， and sugars were the main factors causing the differences in P. ginseng among different forest types. ConclusionThe yield and quality of P. ginseng under Q. mongolica forest were the best， followed by the forest with different tree species， and coming in last was the B. platyphylla forest. This study is expected to provide theoretical support for the improvement of P. ginseng yield and quality and the improvement of ecological planting technology.

Development and Evaluation of Biocontrol Agents Against Panax ginseng Root Rot / 中国实验方剂学杂志

ObjectiveTo develop safe and effective microbial agents against Panax ginseng root rot. MethodP. notoginseng endophytes were screened in plate confrontation tests， followed by morphological and molecular biological identification of antagonistic strains， optimization of strain fermentation conditions in a single factor test， and determination of optimal carriers and auxiliary agents of the microbial agent and their ratio using response surface methodology for formulating the production process. The prevention and control effects of the microbial agent were verified in the confrontation and pot culture experiments. ResultThe plate confrontation test yielded a strain named Fusarium pseudoanthophilum with significant resistance to root rot， and its antibacterial rate was 53.33%. According to the single factor test， the fermentation conditions of F. pseudoanthophilum were determined to be fermentation time 60 h， fermentation temperature 26 ℃， speed 120 r·min-1， and pH 6.5. The response surface optimization results showed that the number of viable bacteria reached the maximum （5.23×109 cfu·g-1） when the peat was 60.00 g， sodium carboxymethylcellulose 3.50 g， and sodium alginate 4.76 g. The influences of carriers and auxiliary agents on the number of viable bacteria were sorted by degree in a descending order as follows： peat>sodium carboxymethylcellulose >sodium alginate. The confrontation test results showed that when the microbial agent concentration was greater than 1.00 g·L-1， it had a significant inhibitory effect on the root rot pathogen F. oxysporum and the inhibitory rate was more than 42.3%. As demonstrated by the pot culture experiment， the inoculation of biocontrol agent for 28 d significantly reduced the incidence （66.99%） of root rot in P. ginseng seedlings and disease index （61.69%） and increased their leaf length （33.04%） and fresh weight （34.48%）. ConclusionF. pseudoanthophilum inoculant is efficient in preventing and controlling the root rot， making it worthy of further development and utilization.

Effects of Exogenous Fusarium oxysporum and Trichoderma viride on Soil Fungal Community of Panax ginseng / 中国实验方剂学杂志

ObjectiveTo investigate the effects of exogenous Fusarium oxysporum and Trichoderma viride on the diversity of soil fungal community and carbon metabolic function of cultivated Panax ginseng. MethodIllumina HiSeq 2500 high-throughput sequencing combined with Biolog-ECO was used to analyze the species diversity and functional diversity of soil fungal communities in P. ginseng soil under different exogenous treatments. ResultThe results of high-throughput sequencing showed that the number and species of microorganisms in the soil were significantly changed after exogenous microorganisms were added. The soil fungi with relative abundance greater than 1% included Mortierella sp.，Fusarium sp.，Humicola sp.，and Simplicillium sp. Mortierella sp. in each treatment group significantly increased. Humicola sp. and Simplicillium sp. could be induced to increase by exogenous addition of F. oxysporum，while T. viride at a high concentration could significantly inhibit the growth of F. oxysporum. As revealed by Biolog and principal component analysis （PCA），the average well color development （AWCD） in the high-dose T. viride group （MG） was significantly higher than that in the control group （QS）and the low-dose F. oxysporum group（LD）. The utilization abilities for amino acids，carboxylic acids，polymers， and amines were enhanced in the MG group，but the microbial metabolic activity was reduced in the high-dose F. oxysporum group （LG）. There was no significant increase in the utilization of phenolic acids by soil microorganisms in both groups. ConclusionExogenous addition of F. oxysporum can lead to the growth and reproduction of other pathogenic fungi. Exogenous addition of T. viride can enhance the soil fungal community structure and metabolic diversity，inhibit the proliferation of F. oxysporum，and improve the soil microbial environment of cultivated P. ginseng.

Research progress on pesticide residues of Panax ginseng / 中国中药杂志

Panax ginseng, a perennial herb, is prone to diseases and insect pests in the growth process, which are primarily prevented and treated by pesticides. However, due to the lack of standardization in the types, frequencies, and doses of pesticides, pesticide residues have become the main exogenous pollutants of P. ginseng. To explore the risk of pesticide residues in P. ginseng, this paper summarized and analyzed the common pesticide residues in P. ginseng, detection techniques, and pesticide residue limit stan-dards based on the published literature in recent years. The results revealed that the main pesticide residues in P. ginseng were organochlorine pesticides, such as tetrachloronitrobenzene, pentachloronitrobenzene, and hexachlorobenzene, and the detection techniques were dominated by gas chromatography(GC), liquid chromatography(LC), or those combined with mass spectrometry(MS). Because of the long half-life and difficulty in degradation, organochlorine pesticides have become the main factor affecting the export of P. ginseng. It is worth mentioning that P. ginseng has been classified as food in Japan, South Korea, the European Union, and other countries, and the standards of pesticide residues and limits are stricter than those in China. The quality and safety of P. ginseng are prerequisites for the efficacy of Chinese medicine and the development of traditional Chinese medicine. The formulation of scientific and effective standards for pesticide application and limits would promote the high-quality development of the P. ginseng industry.

Bioinformatics analysis and function prediction of ginseng TCP transcription factor family / 中国中药杂志

The longevity mechanism of ginseng(Panax ginseng) is related to its strong meristematic ability. In this paper, this study used bioinformatic methods to identify the members of the ginseng TCP gene family in the whole genome and analyzed their sequence characteristics. Then, quantitative real-time fluorescent PCR was performed to analyze the TCP genes containing elements rela-ted to meristem expression in the taproots, fibrous roots, stems, and leaves. According to the data, this study further explored the expression specificity of TCP genes in ginseng tissues, which facilitated the dissection of the longevity mechanism of ginseng. The ginseng TCP members were identified and analyzed using PlantTFDB, ExPASy, MEME, PLANTCARE, TBtools, MEGA and DNAMAN. The results demonstrated that there were 60 TCP gene family members in ginseng, and they could be divided into two classes: Class Ⅰ and Class Ⅱ, in which the Class Ⅱ possessed two subclasses: CYC-TCP and CIN-TCP. The deduced TCP proteins in ginseng had the length of 128-793 aa, the isoelectric point of 4.49-9.84 and the relative molecular mass of 14.2-89.3 kDa. They all contained the basic helix-loop-helix(bHLH) domain. There are a variety of stress response-related cis-acting elements in the promoter regions of ginseng TCP genes, and PgTCP20-PgTCP24 contained the elements associated with meristematic expression. The transcription levels of PgTCP20-PgTCP24 were high in fibrous roots and leaves, but low in stems, indicating the tissue-specific expression of ginseng TCP genes. The Class Ⅰ TCP members which contained PgTCP20-PgTCP23, may be important regulators for the growth and development of ginseng roots.

Panax japonicus and chikusetsusaponins: A review of diverse biological activities and pharmacology mechanism / 中草药·英文版

Panax japonicus, which in the Tujia dialect is known as “Baisan Qi” and “Zhujieshen”, is a classic “qi” drug of Tujia ethnomedicine and it has unique effects on disease caused by “qi” stagnation and blood stasis. This paper serves as the basis of further scientific research and development of Panax japonicus. The pharmacology effects of molecular pharmacology were discussed and summarized. P. japonicus plays an important role on several diseases, such as rheumatic arthritis, cancer, cardiovascular agents, and this review provides new insights into P. japonicus as promising agents to substitute ginseng and notoginseng.

Effects of ecological factors on shape and ginsenoside of Panax ginseng / 中国中药杂志

The ecological environment is closely related to the growth and quality of authentic medicinal materials. Ginseng is very strict with its natural environment and grows mostly in the damp valleys of forests, and the appearance and chemical composition of ginseng under different growth environments are very different. This article reviews the effects of different ecological factors(including light, temperature, altitude, moisture, soil factors, etc.)on the appearance and chemical composition(mainly ginsenosides) of ginseng. Through systematic review, it is found that soil physical factors are the most important ecological factors that affect the appea-rance of ginseng, and soil bulk density plays a key role; temperature affects ginsenosides in ginseng medicinal materials The dominant ecological factors for the accumulation of chemical ingredents; strong light, high altitude, high soil moisture, low soil nutrient and strong acid soil can influence the accumulation of secondary metabolites in ginseng. Environmental stress can also stimulate the formation and accumulation of secondary metabolites in medicinal plants. Appropriate low temperature stress, high or low water stress, acid or alkali stress can also promote the accumulation of ginsenosides. This article systematically reviews the ecological factors that affect the appearance and chemical composition of ginseng, and clarifies the dominant ecological factors and limiting factors for the formation of ginseng's appearance and quality, as well as beneficial environmental stress factors, in order to provide a theoretical basis for ginseng ecological planting and ginseng quality improvement.

Penicillium sp. YJM-2013 induces ginsenosides biosynthesis in Panax ginseng adventitious roots by inducing plant resistance responses / 中草药·英文版

Objective: Fusarium oxysporum is a common pathogenic fungus in ginseng cultivation. Both pathogens and antagonistic fungi have been reported to induce plant resistance responses, thereby promoting the accumulation of secondary metabolites. The purpose of this experiment is to compare the advantages of one of the two fungi, in order to screen out more effective elicitors. The mechanism of fungal elicitor-induced plant resistance response is supplemented. Methods: A gradient dilution and the dural culture were carried out to screen strains. The test strain was identified by morphology and 18 s rDNA. The effect of different concentrations (0, 50, 100, 200, 400 mg/L) of Penicillium sp. YJM-2013 and F. oxysporum on fresh weight and ginsenosides accumulation were tested. Signal molecules transduction, expression of transcription factors and functional genes were investigated to study the induction mechanism of fungal elicitors. Results: Antagonistic fungi of F. oxysporum was identified as Penicillium sp. YJM-2013, which reduced root biomass. The total ginsenosides content of Panax ginseng adventitious roots reached the maximum (48.95 ± 0.97 mg/g) treated with Penicillium sp. YJM-2013 at 200 mg/L, higher than control by 2.59-fold, in which protopanoxadiol-type ginsenosides (PPD) were increased by 4.57 times. Moreover, Penicillium sp. YJM-2013 activated defense signaling molecules, up-regulated the expression of PgWRKY 1, 2, 3, 5, 7, 9 and functional genes in ginsenosides synthesis. Conclusion: Compared with the pathogenic fungi F. oxysporum, antagonistic fungi Penicillium sp. YJM-2013 was more conducive to the accumulation of ginsenosides in P. ginseng adventitious roots. Penicillium sp. YJM-2013 promoted the accumulation of ginsenosides by intensifying the generation of signal molecules, activating the expression of transcription factors and functional genes.

Molecular identification of Sanqi Tablets using DNA barcoding and its methodological evaluation / 中草药

Objective: In order to study the application of DNA barcoding in the authentication of Chinese patent medicines, Sanqi Tablets were used as the object to investigate the applicability, specificity and precision of this method. Methods: Fifteen batches of commercially available Sanqi Tablet samples were collected. The conditions of DNA extraction for Sanqi Tablet had been first investigated, and the DNA was used for testing the applicability of the methods such as PCR amplification, sequence acquisition, and species authentication in the principles for molecular identification of traditional Chinese materia medica using DNA barcoding. The specificity and reproducibility of DNA barcoding in identification of Sanqi Tablets and its adulterations from the roots of Panax notoginseng, P. ginseng and P. quinquefolius were also studied. Results: The Sanqi Tablet sample with an amount of sampling to be 100 mg and a water bath at 56 ℃ for 8 h gave an average concentration of 60.7 ng/μL and then the PCR amplification, sequence acquisition and species assignment were all successful. The ITS2 sequences of P. notoginseng, P. ginseng and P. quinquefolius were all 230 bp in length, and there were seven stable SNP loci between P. notoginseng and P. ginseng, P. notoginseng and P. quinquefolius. ITS2 sequences could be successfully obtained from lab-made and the adulterated Sanqi Tablets, and the Sanger sequencing chromatograms of different ratios of P. notoginseng and P. ginseng mixtures, P. notoginseng and P. quinquefolius mixtures had heterozygous peaks with corresponding peak height ratio at SNP positions. The repeatability, intermediate precision and reproducibility were all in line with the requirements of “General Regulation 9101” in the Chinese Pharmacopoeia. Conclusion: The ITS2 sequence can stably and accurately authenticate the raw materials of Sanqi Tablets with substantial specificity and precision. The DNA barcoding identification method of Sanqi Tablets will provide a new technical tool for ensuring the safety of Sanqi Tablets in clinical medications, and provide reference for the identification of other single-herb products documented in the Chinese Pharmacopoeia.

Expression analysis of transcription factor ERF gene family of Panax ginseng / 中国中药杂志

Ethylene responsive factor(ERF), one of the largest families of transcriptional factors in plants, plays a key role in se-condary metabolism of herbal plants. To analyze the expression of ERF family genes, the heat map clustering method was used by analyzing the ginseng transcriptomes of different parts and different growth years. The contents of ginsenosides Rg_1, Re and Rb_1 in various concentrations of MeJA-treated ginseng adventitious roots were determined by UPLC-MS/MS method. The expression of key genes of ginsenoside biosynthesis(DDS, CYP716A47, CYP716A53v2) and ERF family genes in MeJA-treated ginseng adventitious roots were determined by using real-time quantitative PCR. Pearson correlation was adopted to analyze the gene expression pattern of DDS, CYP716A47, CYP716A53v2 gene and ERF family. The results showed that the content of ginseng diol ginsenoside Rb_1 in ginseng adventitious roots treated with different concentrations of MeJA increased, and the content of ginseng triol ginsenoside Rg_1 and Re decreased. It is consistent with the increase of DDS and CYP716A47 expression and the decrease of CYP716A53v2 gene expression. The expression of ERF003, ERF118 and ERF012 genes was significantly positively correlated with CYP716A53v2, but negatively correlated with DDS. While the expression of ERF1B was significantly negatively correlated with CYP716A47.It is proved that ERF003, ERF118 and ERF012 were likely to inhibit the expression of DDS and promote the expression of CYP716A53v2, and ERF1B was likely to inhibit CYP716A47. This work could provide theoretical basis of ERF functional verification of regulating the biosynthesis of ginsenosides.