A new triterpenoid saponin from the leaves and stems of Panax quinquefolium L.

One new triterpcnoid saponin, quinquenoside L17 （1）, was isolated from the leaves and stems of Panax quinquefolium L., and its structure was elucidated as 20-O-[（β-D-xylopyranosyl-（1-6）-O-β-D-glucopyranosy）]-6-O-β-D-glucopyranosy1-dammar-24-ene- 3,6,12,20-tetraol, by the combination analysis of one-dimensional NMR and two-dimensional NMR, mass spectrometry, CD spectrum and chemical evidences.

Two new acetylated ginsenosides from the roots of Panax quinquefolium

Two new acetylated ginsenosides, 20（S）-protopanaxatriol-6-O-α-L-rhamnopyranosyl （1 → 2）-β-D-6＇-O-acetylglucopyranoside （1） and 20（R）-protopanaxatrol-6-O-α-L-rhamnopyranosyl （1 → 2）-β-o-6＇-O-acetylglucopyranoside （2）, were isolated from the roots of Panax quinquefolium. The structures were elucidated on the basis of spectroscopic techniques and chemical means.

Influence of Panax quinquefolium saponins on increased intracellular Ca^(2+) in PC12 cells

BACKGROUND： Previous studies have demonstrated that intracellular Ca^2＋ （[Ca^2＋]） overload, excitotoxicity, free radical injury, and nitric oxide toxicity are involved in mechanisms of neuronal death in the ischemic brain. OBJECTIVE： To investigate the influence of Panax quinquefo/ium saponins （PQS） on multiple factors-induced Ca^2＋ overload in the rat pheochromocytoma （PC12） cell line. DESIGN, TIME AND SETTING： Intergroup comparison, in vitro study. The experiment was performed at the Heilongjiang Key Laboratory of Anti-fibrosis Biotherapy, Mudanjiang Medical University between November 2007 and April 2008. MATERIALS- In vitro cultured PC12 cells in the logarithmic phase were assigned into blank control, model, and drug treatment groups （10 μmol/L nimodipine; 40 μg/L, 100 μg/L, and 250 μg/L PQS）. Nimodipine was purchased from Jiangsu Yangtze River Pharmacy Group Co., China; PQS （purity 〉 95%, HLPC grade） was provided by School of Basic Medical Sciences, Jilin University. Caffeine, Na2S2O4, L-glutamic acid （Glu）, Fura-2/AM, and calcium ionophore A23187 were purchased from Sigma, USA. METHODS： PC12 cells in the model and drug treatment groups were separately incubated in glucose-free Hank＇s buffered saline solution ＋ Na2S2O4 （2 mmol/L） for 6 hours, Glu （200 μmot/L） plus A23187 （0.05 μmol/L） for 6 hours, KCI （50 mmol/L） for 1 hour, and caffeine （5 mmol/L） for 3 hours to establish models of intracellular Ca^2＋ overload induced by oxygen and glucose deprivation, Glu, A23187, high K＋, or caffeine. In addition, control cells were incubated in high-glucose DMEM culture medium. MAIN OUTCOME MEASURES： [Ca^2＋]i changes in PC12 cells exposed to oxygen-glucose deprivation, Glu, A23187, high K^＋, or caffeine were detected using spectrofluorometer. RESULTS： PQS blocked the [Ca^2＋]i increase induced by oxygen-glucose deprivation, Glu, A23187, high K＋, or caffeine. In particular, high-dose PQS was most effective （P 〈 0.01）. PQS significantly inhibited Glu- or caffeine-induced [Ca^2＋]i increases in the absence of extracellular Ca^2＋, but nimodipine did not. CONCLUSION： PQS blocked intracellular Ca^2＋ overload induced by oxygen-glucose deprivation, Glu, A23187, high K^＋, or caffeine. This mechanism might be involved in the attenuation of neuronal apoptosis following ischemic brain injury.

A new ceramide from transgenic crown galls of Panax quinquefolium

A new ceramide （1） was isolated from transgenic crown galls of Panax quinquefolium. The structure was elucidated as （2S, 3S, 4R, 20E）-2-[（2＇R）-2＇-hydroxylpalmitoylamino]-20-hexacosene- 1, 3, 4-triol on the basis of spectroscopic and chemical methods.

Isolation and Characterization of a Bioactive Polysaccharide from Panax Quinquefolium L.

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Research Progress on Antitumor Effect and Mechanism of Panax quinquefolium Linn.

Panax quinquefolium Linn.,belonging to Panax,Araliaceae,contains a variety of active ingredients.In recent years,many studies have shown that P.quinquefolium Linn.has high value and development prospect in inhibiting colon cancer,lung cancer and breast cancer.Recent advances in studies on chemical components,antitumor effects and mechanisms of P.quinquefolium Linn.are reviewed in the paper.

基于转录组学测序探讨西洋参、丹参配伍对心肌梗死后大鼠心肌缺血的影响

目的:运用转录组学测序探讨西洋参、丹参治疗心肌梗死后心肌缺血的作用靶点。方法:通过结扎左冠状动脉前降支构建心肌梗死大鼠模型。将造模成功的大鼠随机分为模型组(MOD组,等量蒸馏水)、西洋参+丹参低剂量组[XD-L组,西洋参1.5 g/(kg·d)+丹参4.5 g/(kg·d)]、西洋参+丹参高剂量组[XD-H组,西洋参3 g/(kg·d)+丹参9 g/(kg·d)]、ACEI组[培哚普利0.84 mg/(kg·d)];并以假手术大鼠作为对照组(CON组,等量蒸馏水),每组6只,连续灌胃4周。采用超声心动图测定大鼠心功能;苏木精-伊红(HE)染色观察大鼠心肌组织病理改变。采用转录组学测序筛选西洋参、丹参治疗心肌梗死后缺血区的差异表达基因(DEGs),并对DEGs进行基因本体(GO)、京都基因和基因组百科全书(KEGG)富集分析。结果:西洋参、丹参可改善心肌梗死后大鼠心功能,减轻心肌组织病理损伤。转录组学测序结果表明,与CON组比较,MOD组大鼠12个基因上调,13个基因下调。西洋参、丹参回调了MOD组的6个基因。DEGs主要富集在三羧酸(TCA)循环、磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/AKT)等信号通路;与免疫炎症反应、细胞凋亡、血管新生等有关。结论:西洋参、丹参可能通过上调Wnt4表达,抑制细胞凋亡和炎症反应,从而改善心肌梗死后大鼠心功能。

三七与西洋参种间杂交F1代无性系SSR分子标记及倍性鉴定

目的:通过SSR分子标记技术及流式细胞术对三七Panax notoginseng(Burk.)F.H.Chen与西洋参Panax quinquefolium L.杂交F1代杂种进行真实性鉴定。方法:设计、筛选引物,对随机选取的60份杂交F1代无性系幼苗进行SSR分子鉴定;裂解样品细胞,利用流式细胞仪对60份样品进行倍性分析。结果:筛选出Q2、Q5两条引物可用于三七与西洋参杂交F1代的鉴定,在60份样品中,共有39份具有双亲互补条带;利用流式细胞仪对相同的60份样品进行检测,三倍体样品数为39份,表现为双亲互补条带的样品倍性检测结果均为三倍体,可认定为真杂种。结论:杂交F1代的真实杂种率为65%,SSR分析与倍性检测结果相互印证,证实了鉴定结果的可靠性。

西洋参加工副产物对黄羽肉鸡血清生化指标、抗氧化能力和肉品质的影响

为研究西洋参加工副产物对黄羽肉鸡血清生化指标、抗氧化能力及肉品质的影响。本试验选择7日龄黄羽肉鸡160只,随机分成4组,每组4个重复,Ⅰ组、Ⅱ组、Ⅲ组分别添加1%、3%、5%西洋参加工副产物,Ⅳ组为基础饲粮对照组,试验期100d。结果表明,西洋参加工副产物可显著降低肉鸡血糖含量(P <0.05),并显著降低血清中低密度脂蛋白含量(P <0.05);试验组总抗氧化力和谷胱甘肽过氧化物酶极显著高于对照组(P <0.01);Ⅰ组和Ⅱ组的胸肌、腿肌pH值、亮度、红度和黄度极显著优于对照组(P<0.01);试验组胸肌滴水损失和剪切力显著优于对照组(P <0.05)。在本试验条件下,饲粮中添加1%~3%的西洋参加工副产物,能够较好的调节胸肌、腿肌外观色泽,更符合市场消费的需求。

西洋参根腐病拮抗菌XT-25的生防作用研究

为探讨西洋参根腐病的生物防治技术,本研究以西洋参根腐病病原茄病镰孢菌(Fusarium solani)和土赤壳属的Ilyonectria mors-panacis为靶标,从西洋参根际土中筛选具有拮抗作用的生防菌,通过对峙培养、菌丝抑制活性、抑菌促生物质测定和离体接种防效等试验,初步探讨拮抗菌株对西洋参根腐病的抑菌机制和防治效果。结果表明,分离到1株对2种西洋参致病菌的抑菌率皆在55%以上的菌株XT-25;结合形态学特征及16S rRNA、atpD和rpoB基因序列系统发育分析结果,将菌株XT-25鉴定为淡紫灰链霉菌(Streptomyces lavendulae);菌株XT-25对黄瓜枯萎病菌等6种病原菌的抑菌率为59%~70%。菌株XT-25不仅具有分泌蛋白酶、淀粉酶以及β-1,3葡聚糖酶等抑菌物质的能力,还具有溶解有机磷、无机磷、产氨、固氮、以及分泌IAA和ACC脱氨酶等促生物质能力。菌株XT-25挥发性气体对F.solani的抑菌率为9.04%;其可造成F.solani、I.mors-panacis菌丝的畸形。离体接种西洋参的防效试验结果表明,菌株XT-25对致病菌F.solani和I.mors-panacis引致的西洋参根腐病的防治效果分别为40.00%和24.74%。本研究结果为防治由镰孢菌和土赤壳菌引起的西洋参根腐病提供了生防材料和理论参考。

Panax quinquefolium saponin Optimizes Energy Homeostasis by Modulating AMPK-Activated Metabolic Pathways in Hypoxia-Reperfusion Induced Cardiomyocytes

Objective To investigate the effects and underlying mechanisms of Panax quinquefolium saponin(PQS)on energy deficiency in hypoxia-reperfusion(H/R)induced cardiomyocytes.Methods The H/R injury involved hypoxia for 3 h and then reperfusion for 2 h.Cardiomyocytes recruited from neonatal rat ventricular myocytes(NRVMs)were randomly divided into control,H/R,H/R+compound C(C.C),H/R+PQS,and H/R+C.C+PQS groups.BrdU assay,lactase dehydrogenase(LDH)leakage and early apoptosis rate were evaluated to assess cell damages.Contents of high energy phosphate compounds were conducted to detect the energy production.Protein expression levels of adenosine monophosphate-activated protein kinase a(AMPKα),glucose transporter 4(GLUT4),phosphate fructose kinase 2(PFK2),fatty acid translocase/cluster of differentiation 36(FAT/CD36),and acetyl CoA carboxylase 2(ACC2)in the regulatory pathways were measured by Western blotting.Immunofluorescence staining of GLUT4 and FAT/CD36 was used to observe the mobilization of metabolic transporters.Results PQS(50 mg/L)pretreatment significantly alleviated H/R-induced inhibition of NRVMs viability,up-regulation of LDH leakage,acceleration of early apoptosis,and reduction of energy production(P<0.05).Compared with the H/R group,up-regulated expression of AMPKα,GLUT4,PFK2,FAT/CD36 and ACC2 were observed,and more GLUT4 and FAT/CD36 expressions were detected on the membrane in the H/R+PQS group(P<0.05).These effects of PQS on H/R-induced NRVMs were eliminated in the H/R+C.C+PQS group(P<0.05).Conclusion PQS has prominent advantages in protecting NRVMs from H/R-induced cell damages and energy metabolic disorders,by activation of AMPKα-mediated GLUT4-PFK2 and FAT/CD36-ACC2 pathways.

Effects of Panax Quinquefolium Saponin on Phosphatidylinositol 3-Kinase/Serine Threonine Kinase Pathway of Neonatal Rat Myocardial Cells Subjected to Hypoxia

Objective： To explore the effects of Panax Quinquefolium Saponin （PQS） on phosphatidylinositol 3-kinase/serine threonine kinase （P13/Akt） pathway of neonatal rat myocardial cells subjected to hypoxia. Methods： Neonatal rat myocardial cells were cultured in vitro. After the myocardial cell injury was induced by hypoxia, the cells were randomized into 5 groups： the normal group, the model group, the positive control group （Ciclosporin A, 2 p, mol/L）, the low-dose PQS group （PQSL, 25mg/L）, and the high-dose PQS group （PQSH, 50 mg/L）. Morphology and behavior of myocardial cells were observed under an inverted microscope. Apoptosis rate and lactate dehydrogenase （LDH） leakage rate of myocardial cells were determined by colorimetry. Mitochondrial transmembrane potential was assessed using a fluorexon laser. Phospho-glycogen synthase kinase （GSK）-3β and phospho-Akt as well as cytochrome C were determined by Western blot. Results： LDH leakage in the Ciclosporin A group, PQSH group and PQSL group reduced progressively compared with the model group （P〈0.05）. Akt and GSK-3β was strongly phosphorylated after treatment with Ciclosporin A and PQS compared with the model group （P〈0.05, P〈0.01）. Compared with the model group （16.41 ± 1.74; 35.28 ± 6.30）, both the integrated optical density of mitochondrial permeability transition pore （MPTP） and the mitochondrial transmembrane potential significantly increased in the PQSH group （42.74± 2.12; 71.36 ± 6.54） and the PQSL group （39.58± 1.49; 66.99± 5.45; P〈0.05, P〈0.01）. However, the protein of cytochrome C outside the mitochonddon decreased in the PQSH group （273.66 ± 14.61） and the PQSL group （259.62 ± 17.31） compared with the model group （502.41 ± 17.76; P〈0.05）. Conclusion： Through activation of the P13K/Akt pathway and inhibition of the MPTP, PQS might protect the heart against ischemia injury and apoptosis of myocardial cells.

UPLC-MS/MS测定心悦胶囊中6个皂苷类成分的含量

目的:采用超高效液相色谱-质谱法(UPLC-MS/MS)建立心悦胶囊中主要成分人参皂苷Rg_(1)、人参皂苷Re、人参皂苷Rb_(1)、拟人参皂苷F_(11)、人参皂苷Rd和20(S)-人参皂苷F1的含量测定方法。方法:使用Waters ACQUITY BEH C_(18)色谱柱(100 mm×2.1 mm,1.7μm),以乙腈-水为流动相,梯度洗脱,流速为0.5 mL·min^(–1),柱温为40℃,电喷雾电离源,多反应离子监测扫描方式进行检测。结果:6个成分分别在质量浓度为9.638~963.800、9.839~983.900、9.951~995.100、5.270~1054.000、9.608~960.800、4.905~981.000 ng·mL^(–1)时与峰面积线性关系良好(r>0.998),平均加样回收率(RSD)分别为96.9%(2.2%)、100.1%(1.5%)、97.2%(1.8%)、98.5%(2.3%)、96.6%(2.8%)、100.2%(3.5%)。结论:所建立的方法快速、准确、灵敏度高,可用于心悦胶囊中西洋参茎叶总皂苷的质量控制。

Integrative SMRT sequencing and ginsenoside profiling analysis provide insights into the biosynthesis of ginsenoside in Panax quinquefolium

Panax quinquefolium is one of the most common medicinal plants worldwide.Ginsenosides are the major pharmaceutical components in P.quinquefolium.The biosynthesis of ginsenosides in different tissues of P.quinquefolium remained largely unknown.In the current study,an integrative method of transcriptome and metabolome analysis was used to elucidate the ginsenosides biosynthesis pathways in different tissues of P.quinquefolium.Herein,22 ginsenosides in roots,leaves,and flower buds showed uneven distribution patterns.A comprehensive P.quinquefolium transcriptome was generated through single molecular real-time(SMRT)and second-generation sequencing(NGS)technologies,which revealed the ginsenoside pathway genes and UDP-glycosyltransferases(UGT)family genes explicitly expressed in roots,leaves,and flower buds.The weighted gene co-expression network analysis(WGCNA)of ginsenoside biosynthesis genes,UGT genes and ginsenoside contents indicated that three UGT genes were positively correlated to pseudoginsenoside F11,notoginsenoside R1,notoginsenoside R2 and pseudoginsenoside RT5.These results provide insights into ginsenoside biosynthesis in different tissues ofP.quinquefolium.

UPLC法测定西洋参不同部位中皂苷类成分及质量评价研究

本文旨在建立超高效液相色谱-二极管阵列检测器(UPLC-PDA)法同时测定山东西洋参中16种皂苷类成分的方法,并对山东省威海市5个地区11批西洋参芦头、主根、侧根和须根不同部位的皂苷含量进行测定,以期为山东西洋参质量控制与评价提供科学依据。采用Acquity UPLC BEH C_(18)柱(2.1 mm×50 mm,1.7μm),流动相为水-乙腈,梯度洗脱,流速0.4 mL/min,柱温为30℃,进样量为2μL,PDA检测器,检测波长为203 nm。16种皂苷在0.05~1.00 mg/mL范围内线性良好,相关系数均大于0.999 1,加标回收率为92.45%~103.24%,RSD为0.31%~4.60%,灵敏度、准确度及精密度均符合要求。山东省威海地区西洋参芦头、主根、侧根和须根部位皂苷含量分别为(42.69±7.01) mg/g、(32.63±4.71) mg/g、(30.80±5.15) mg/g和(37.55±5.23) mg/g。通过主成分分析及正交偏最小二乘法分析,初步筛选出人参皂苷Re、Rc、Rb_(1)、Rb_(3)和Rb_(2)作为西洋参不同部位鉴别的潜在质量标志物。本研究建立的方法可用于西洋参皂苷含量测定及不同部位的区分,对西洋参质量进行初步评价。

Purification and Identification of a Novel Protein Isolated from Panax quinquefolium and Evaluation of its In vitro Antioxidant Properties

Objective:A novel protein was first purified from Panax quinquefolius L.(AGNP),and in vitro antioxidant activities of AGNP were first studied in this work.Methods:AGNP was purified by Ion-exchange chromatography and Gel-filtration chromatography.The chemical characterizations of AGNP were tested by sodium dodecyl sulfate–polyacrylamide gel electrophoresis(SDS-PAGE),high-pressure gel-filtration chromatography,MALDI-TOF-MS and HPLC.In vitro antioxidant effects were tested in simple antioxidant assay including 2,2-diphenylpicrylhydrazyl radical scavenging,superoxide radical(O2^-)scavenging,hydroxyl radical(OH)scavenging,and ferric-reducing ability.Results:The results showed which the content of AGNP measured by Bradford method was 2.42 mg/m L and the subunit molecular weight of AGNP measured by sodium dodecyl sulfate–polyacrylamide gel electrophoresis(SDS-PAGE)was 15 kD.The AGNP molecular weight was 15,114 Da both of SDS-PAGE and mass spectrum purity.The result of high-pressure gel-filtration chromatography demonstrated that the molecular weight of AGNP was 31,086 Da,which implied that AGNP was a homodimer.The in vitro Antioxidant results indicated that AGNP had obvious effects to remove the free radicals in vitro.Conclusion:In conclusion,AGNP had more powerful antioxidant capacity and it can be used as an effective natural antioxidant to alleviate oxidative stress.

西洋参茎叶皂苷对力竭运动大鼠心肌损伤的改善作用

为了研究西洋参茎叶皂苷减轻力竭运动大鼠心肌损伤的作用及机制,为西洋参茎叶皂苷的综合利用提供参考,本试验分为正常对照组、力竭运动组及西洋参茎叶皂苷低、中、高剂量组(100、200、400 mg/kg),除正常对照组外均需进行力竭游泳训练,4周后检测相关指标变化。结果表明,(1)与力竭运动组相比,西洋参茎叶皂苷中、高剂量组大鼠LVIDs、LVIDd极显著减少(P<0.01),低、中、高剂量组LVEF、LVFS极显著增加(P<0.01),心肌组织cTnⅠ含量及血清CTNK、LDH、ALT、α-HBDH活性均极显著降低(P<0.01);中、高剂量组LVIDs、LVIDd同低剂量组比极显著减少(P<0.01),高剂量组LVEF极显著高于低、中剂量组(P<0.01),中、高剂量组LVFS极显著高于低剂量组(P<0.01),而CTNK、α-HBDH活性极显著低于低剂量组(P<0.01),高剂量组LDH活性极显著低于低、中剂量组(P<0.01),低、中、高剂量组ALT活性比较差异均有极显著性(P<0.01)。(2)与力竭运动组相比,西洋参茎叶皂苷低、中、高剂量组大鼠心肌组织及血清MDA含量极显著降低(P<0.01),除低剂量组大鼠血清GSH-Px活性显著增加(P<0.05)外,其余各组大鼠心肌组织及血清SOD、CAT及GSH-Px活性均极显著增加(P<0.01);高剂量组心肌组织及血清MDA含量极显著低于低、中剂量组(P<0.01),SOD、CAT活性则极显著高于低、中剂量组(P<0.01),中、高剂量组GSH-Px活性极显著高于低剂量组(P<0.01)。(3)与力竭运动组相比,西洋参茎叶皂苷低剂量组大鼠心肌组织细胞凋亡率显著降低(P<0.05),中、高剂量组细胞凋亡率及各剂量组Bax、cleaved caspase-3表达极显著降低(P<0.01),而Bcl-2表达极显著增加(P<0.01);中、高剂量组细胞凋亡率、Bax及cleaved caspase-3表达均极显著少于低剂量组(P<0.01),而Bcl-2表达极显著高于低剂量组(P<0.01)。本研究显示,西洋参茎叶皂苷具有减轻力竭运动大鼠心肌损伤及改善心功能作用,且该作用与抑制氧化应激及抗心肌细胞凋亡有关。

UPLC-MS/MS检测补肾益脑胶囊中人参掺伪情况

目的:建立一种可测定补肾益脑胶囊制剂中非法添加西洋参的超高效液相色谱-质谱法,考察企业是否存在使用西洋参或其边角料替代人参投料的现象,为中药监管提供技术支持。方法:Phenomenex C_(18)色谱柱(100 mm×2.1 mm,2.6μm),以10 mmoL·L^(-1)的乙酸铵(A)-乙腈(B)为流动相梯度洗脱;流速为0.3 mL·min^(-1);柱温为40℃。采用电喷雾负离子模式进行多反应监测。结果:60批补肾益脑胶囊中有29批检出西洋参的专属成分拟人参皂苷F_(11),检出率为48.3%。结论:补肾益脑胶囊中存在人参掺伪的现象,该方法准确可靠,可作为补肾益脑胶囊中人参掺混西洋参的补充检验方法。

基于离子液体原位泡腾萃取法在丹参及西洋参克百威残留分析中的应用

本研究建立了一种新颖的基于酸性离子液体的原位泡腾萃取技术的前处理方法。酸性离子液体1-丁基-3-甲基咪唑硫酸氢盐呈酸性,能与碳酸氢盐反应产生二氧化碳,起到泡腾萃取的效果,它还能与六氟磷酸铵发生离子交换作用而置换成疏水性离子液体1-丁基-3-甲基咪唑六氟磷酸盐,使其与水相分离,通过离心获取萃取相,最后通过高效液相色谱仪测定萃取相中农药的含量。考察了酸性离子液体、碳酸氢钠和氯化钠用量对萃取效率的影响,最终所得最佳条件为200 mg酸性离子液体、0.4 g碳酸氢钠和1.0 g氯化钠。在最优条件下,克百威的线性范围为0.3~4.0μg/mL,日内和日间的相对偏差(RSD)为4.6%和10.2%,丹参和西洋参在4.0μg/mL加标水平下,加标回收率为94.7%(西洋参)和74.6%(丹参),将此方法可用于不同基质中药材和饮片中农药残留的处理。

Microbial inoculants and garbage fermentation liquid reduced root-knot nematode disease and As uptake in Panax quinquefolium cultivation by modulating rhizosphere microbiota community

Objective:To find a suitable ecological cultivation measure to solve the problem of root-knot nematode disease of Panax quinquefolium(Panacis Quinquefolii Radix)and the heavy metals accumulating in its roots.Methods:Three-year-old P.quinquefolium was treated with four different combinations of microbial inoculant(MI)and garbage fermentation liquid(GFL)[the joint application of‘TuXiu’MI and Fifty potassium MI(TF),the combination use of‘No.1'MI and Fifty potassium MI(NF),‘Gulefeng’poly-γ-glutamic acid MI(PGA),GFL],and the untreated control(CK).Here,high-throughput sequencing,ICP-MS and UPLC were employed to systematically characterize changes of microbial diversity and structure composition,heavy metals(As,Cd and Pb)content and ginsenoside content among different treatments.Results:The results revealed that different MIs and GFL could increase the root dry weight of P.quinquefolium,PGA enhanced it by 83.24%,followed by GFL(49.93%),meanwhile,PGA and GFL were able to lessen root-knot nematode disease incidence by 57.25%and 64.35%.The treatment of PGA and GFL can also effectively reduce heavy metals in roots.The As content in GFL and PGA was decreased by 52.17%and 43.48%respectively,while the Cd and Pb contents of GFL and PGA was decreased somewhat.Additionally,the content of total ginsenosides was increased by 42.14%and 42.07%,in response to TF and NF,respectively.Our metagenomic analysis showed that the relative abundance of particular soil microbial community members related to the biocontrol of root-knot nematode disease and plant pathogen(i.e.,Chaetomium in NF,Xylari in GFL,and Microascus in PGA),heavy metal bioremediation(Hyphomacrobium in PGA and Xylaria in GFL),and nitrogen fixation(Nordella and Nitrospira in TF)was significantly increased;notably,potential harmful microflora,such as Plectosaphaerella and Rhizobacter,were more abundant in the control group.Conclusion:MI and GFL could improve the quality of P.quinquefolium by modifying its rhizosphere microbial community structure and composition,both of them are beneficial to the development of ecological cultivation of P.quinquefolium.