电子束辐照对人参抗肿瘤作用的影响

为了明确电子束辐照对人参抗肿瘤作用的影响,以长白山林下参为研究对象,电子束辐照后测定人参皂苷的含量,Hek-293、HCT-116、Hela、HepG2细胞的抑制情况以及荷瘤小鼠的治疗情况,并测定荷瘤小鼠血清MDA含量、T-SOD活性和T-AOC水平。结果表明,辐照能够提高人参中皂苷的含量,其中人参皂苷Rg3含量变化明显。体外试验中,30 kGy辐照组人参提取物对HCT-116、Hela、Hep G2细胞的最高抑制率分别达到75.63%、85.33%、84.87%,均显著高于未辐照人参组;体内试验中,辐照人参组瘤重低于阴性对照组,同时能够提高荷瘤小鼠体内抗氧化能力,从而提高小鼠机体抗肿瘤能力。本试验结果为人参抗肿瘤作用的研究提供了新的思路。     

一株人参根区解磷细菌的筛选、鉴定及对人参生长的影响

人参是我国传统名贵中药材,也是吉林省对外交往中的代名词。吉林参畦土壤有效磷含量低,但大量施用磷肥又会导致人参品质的下降,因此,本研究从人参根区土壤中筛选高效解磷细菌,接种到人参根部,研究其对人参生长、品质、土壤速效养分、土壤酶活性的影响,以期为人参解磷微生物肥料提供优良菌株,从而促进人参产业绿色发展。采用PKO平板初筛及摇瓶复筛从野山参根区土壤筛选高效解磷细菌。盆栽试验以3年生人参种苗为材料,每盆种植6棵单株重相同的人参种苗,设对照(CK)和处理组,待完全展叶后间苗,留取4棵长势一致的人参苗,采用滴灌的方式将菌液接种在人参苗根部附近,生长至红果期(90 d)取样,测定植物生长量和叶绿素含量、土壤速效养分、参根和参叶中NPK的含量、土壤酶活性、参根中微量元素和8种皂苷的含量。结果表明,1)平板初筛和摇瓶复筛得到P1、P7菌株,摇瓶培养中,P1可溶性磷为472.85μg/m L,P7可溶性磷为437.33μg/m L。2)P1处理的人参参根鲜重、干重显著高于P7、BM、CK处理,株高显著高于对照,叶绿素含量显著高于P7、BM、CK(P0.05)。3)P1处理促进人参对土壤钾元素的吸收,提高磷素在参根中的积累量。4)接种菌剂可以不同程度的提高土壤酶活性,以P1处理的效果最显著,P1处理人参根际土壤脲酶、酸性磷酸酶、过氧化物酶活性均为最高。5)P1处理显著提高了人参根中Fe、Mn、Cu的含量,分别较对照增加了52.74%、127.03%、28.96%,P1处理Zn与对照相比增加19.96%;人参根中8种单体皂苷P1处理均显著高于P7、BM、CK。6)经16S rRNA分析结合P1形态初步鉴定为假单胞菌。     

铜胁迫对人参生理参数和品质的影响研究

为探讨铜胁迫对人参(Panax ginseng C. A. Mey)生理参数及品质的影响机制,以三年生人参种苗为材料,水培处理21 d后研究铜对人参幼苗生长、各部位及对应细胞壁铜含量、生理特性及总皂苷含量的影响。结果表明,高浓度铜胁迫抑制人参生长,破坏细胞正常结构,干重鲜重较对照显著下降。随着铜处理浓度增大,人参各部位铜离子含量显著上升,根、茎、叶最高铜离子含量达48.30、68.58、130.77 mg·kg^-1,铜离子主要分布在细胞壁内,根、茎、叶细胞壁所含铜量占总铜量均在150μmol·L^-1 CuSO₄处理时最高,分别为57.74%、43.33%和37.24%。木质素含量随铜胁迫浓度增加而上升,相关合成酶活性也显著增强,在500μmol·L^-1 CuSO₄处理时达到最大值。人参根和叶H₂O₂及丙二醛(MDA)含量随铜处理浓度增大而上升;过氧化物酶(POD)和过氧化氢酶(CAT)活性在300μmol·L^-1...     

水杨酸诱导下人参培养物差异表达基因片段的克隆

本研究以人参愈伤悬浮细胞为材料,在其生长的第28天添加1×10^-3mg／L水杨酸,测定水杨酸添加后,过氧化物酶、多酚氧化酶和苯丙氨酸解氨酶等3种酶在72h内的变化及皂苷含量,结果表明：水杨酸添加后对过氧化物酶和苯丙氨酸解氨酶的活力影响最大,分别在24h和48h达到最大峰值,在18h开始影响多酚氧化酶的活力,培养物生长的第28天添加水杨酸可以明显提高人参愈伤组织中皂苷的合成。确定添加水杨酸后24h提取总RNA,进行cDNA-RDA分析,筛选差异片段。确定差异基因并在GenBank中注册,注册号为FE900130。为探讨水杨酸作为诱导子对人参次生代谢的影响奠定基础。     

表面活性剂对超临界CO_2萃取人参中皂苷的影响

为了改善超临界CO2萃取在极性物质方面存在的局限性,在其体系中引入特定表面活性剂和助表面活性剂,考察了它们对超临界CO2萃取人参中皂苷的影响。试验结果发现,表面活性剂和助表面活性剂的加入均可显著提高超临界CO2萃取人参中皂苷的萃取率,其改善效果与它们的种类和加入量有关。在司盘80、吐温80、聚乙二醇辛基苯基醚和琥珀酸二(2-乙基己基)酯磺酸钠4种表面活性剂中,以琥珀酸二(2-乙基己基)酯磺酸钠的改善效果最好,其次是聚乙二醇辛基苯基醚和吐温80,而司盘80最差。在3种助表面活性剂对琥珀酸二(2-乙基己基)酯磺酸钠/超临界CO2反相微乳萃取人参皂苷的改善效果方面,以乙醇效果最好,其次是正戊醇,正丁醇效果最差。在萃取压力32MPa、萃取温度45℃、萃取时间4h和CO2流量2.5L/h的条件下,AOT和乙醇的加入量以0.036g/mL较好,此时人参皂苷的萃取率达15.9%,是没加表面活性剂和助表面活性剂下的13.3倍。     

椰糠复配基质在人参穴盘育苗中的应用

为探究椰糠替代草炭用作人参穴盘育苗基质的可行性,将椰糠、草炭、蛭石按体积进行不同比例复配,研究不同配比基质对人参种苗生长及生理指标的影响。结果表明:随椰糠在基质中占比的减少,基质通气孔隙度上升,持水孔隙度下降,有效磷、速效钾含量显著高于CK(草炭 ∶ 蛭石=5∶3),其中T3处理(椰糠 ∶ 蛭石=2∶1)在茎粗、根长、鲜重、干重、根冠比、壮苗指数、叶绿素含量、过氧化物酶(POD)活性、过氧化氢酶(CAT)活性、丙二醛(MDA)含量与CK相比无显著差异,尤以T4处理(椰糠 ∶ 蛭石=1∶1)最优,叶面积、地下干重和鲜重、根冠比、壮苗指数、可溶性蛋白、SOD活性、POD活性、MDA含量显著优于CK(P<0.05),隶属函数综合分析得分(0.936)高于CK(0.775)。综上所述,T4处理为最适的椰糠配比基质,能够替代以草炭为主的人参育苗基质配方。     

不同生育期同化的光合产物在人参体内的转移和分配

在人参不同生育期,采用(14)~CO_2标记整体植株的方法,测定人参同化(14)~C产物的分配比率和残留率。结果表明,展叶末期同化的光合产物60％用于建造人参营养体,为地上部各器官的生长发育提供碳源和能源;绿果期以后的同化产物是参根中干物质和有效成份构成的主要来源;施用人参复合肥料,4年生参根干重及总皂甙含量较对照提高16.1％和4.3％。     

基于产量品质及水肥利用效率的三七水肥耦合方案优选

为了实现三七增产、提质和绿色生产,探究适宜的水肥耦合方案,以2年生三七为研究对象进行大田试验,设置3种灌水水平,分别为低水W1（0.5FC,FC为田间持水量）、中水W2（0.7FC）、高水W3（0.9FC）;4种施肥水平,全年施肥总量为1 440 kg/hm2,根据各个生育期设置不同的施肥比例,划分为F1（苗期∶花期∶果期∶根增重期=25%∶25%∶25%∶25%）、F2（25%∶30%∶25%∶20%）、F3（30%∶30%∶25%∶15%）和F4（40%∶20%∶30%∶10%）,共12种处理,分析水肥及其耦合效应,对三七产量、品质、水分利用效率（Water Use Efficiency,WUE）和肥料偏生产力（Partial Factor Productivity of fertilizer,PFP）的影响,应用优劣解距离法（TOPSIS）法对三七皂苷含量进行综合品质评价,并运用综合评分法对各处理进行基于产量、品质、水肥利用效率的综合评价,探究三七生长与生产管理的综合最优处理。结果表明：水肥耦合对发病率影响最大,灌水对WUE影响最大,生育期不同比例施肥对产量和PFP影响最大;5种皂苷中,W2F4处理的三七皂苷R1、人参皂苷Rg1、人参皂苷Re和人参皂苷Rd的积累量都显著（P＜0.05）高于其他处理,W2F4处理的三七皂苷R1含量最高,W1F4处理的人参皂苷Rb1含量最高;通过综合评分法求得最优处理为W2F4处理,发病率为12.97%、产量为2 976.42 kg/hm2、WUE为1.65 kg/m3、PFP为1.09 kg/kg;TOPSIS品质分析W2F4处理得分最高,为0.815。该研究结果可为三七合理水肥管理提供科学依据。     

三种人参属植物的EST-SSR信息分析及其在三七中的应用

为了解人参属植物EST中SSR分布特点及其在三七SSR标记中的应用,利用生物信息学方法,用primer3软件对dbEST数据库中人参属植物人参、西洋参和三七的EST序列进行搜索,发现人参属植物EST-SSR出现频率为11.54%,平均每4.39kb出现1个SSR。人参属植物单核苷酸重复基元占主导地位,其次为三核苷酸重复基元和二核苷酸重复基元,分别占总SSR的54.48%、17.31%和16.36%。人参属EST-SSR的优势类型为A/T和AT/TA,分别占54.73%和8.21%。根据人参属植物EST中的SSR设计48对引物,在合适的PCR扩增体系下,用5个三七样品的DNA为模板进行PCR扩增,引物有效扩增率85.42%,其中多态性引物占可扩增引物的70.73%。结果表明,利用人参属EST序列开发三七EST-SSR标记是可行的。     

不同参龄人参及其根际土壤中Cu、Zn、Mn含量研究

通过对不同参龄人参药材与根际土壤中Cu、Zn、Mn3种微量元素动态变化特征的分析,探讨人参与土壤中微量元素间的相互关系,找出人参对Cu、Zn、Mn的吸收积累规律和参地土壤Cu、Zn、Mn含量的变化规律,为确定人参适宜的生产环境和建立标准化的田间管理提供依据,结果表明,人参根际土壤中Cu含量随人参种植年限的增加有明显的累积趋势,该人参基地土壤Zn含量偏低,应适当施用Zn肥来提高土壤Zn含量;7月份是各参龄人参药材Cu含量和单株根Cu积累量增加最快的时期,7、8月份是各参龄人参药材Mn含量和单株根Mn积累量增加最快的时期,9月份是6年生人参对Zn元素需求旺盛的时期,在整个人参生长期,6年生人参药材Cu、Mn、Zn含量和单株根积累量都高于4、5年生人参,在本试验的采样年限内,以生长6年为最佳采收年限。     

Simultaneous determination of ginsenosides and polyacetylenes in American ginseng root (Panax quinquefolium L.) by high-performance liquid chromatography

A method for simultaneous determination of ginsenosides and polyacetylenes in Panax quinquefolium L. (American ginseng) roots was developed. The ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Ro, malonyl-Rb1, malonyl-Rc, and malonyl-Rd and the polyacetylenes falcarinol and panaxydol were extracted from fresh ginseng roots in a sequential extraction process with 100% methanol followed by 80% aqueous methanol and quantified simultaneously in extracts by high-performance liquid chromatography using diode array detection. Separations were achieved with a phosphate buffer-acetonitrile gradient system using an RP-C18 column. Except for Rd, the present extraction method resulted in similar or significantly higher concentrations of both ginsenosides and polyacetylenes in comparison to commonly used extraction methods for these compounds. The contents of polyacetylenes and ginsenosides were determined in the root hairs, lateral roots, and main roots of 6 year old ginseng plants. The total mean concentrations of ginsenosides and polyacetylenes in root hairs were 31.0 g/kg fresh weight (FW) and 2.6 g/kg FW, respectively, whereas the concentrations of these bioactive compounds in the main roots were significantly lower with total mean concentrations of 17.8 g/kg FW for ginsenosides and 0.6 g/kg FW for polyacetylenes. The concentration of individual and total ginsenosides and polyacetylenes did not differ significantly between main roots of different sizes. Consequently, it is possible to do quantitative screening for ginsenosides and polyacetylenes to breed ginseng roots with higher levels of bioactive compounds.     

Phytochemistry of wild populations of Panax quinquefolius L. (North American ginseng)

A survey of the phytochemistry of Panax quinquefolius L. (North American ginseng) collected from wild populations in Ontario, Quebec, Maine, Vermont, and Wisconsin was undertaken. Reverse-phase HPLC was used to determine the natural variation of levels of ginsenosides Rg1, Re, Rf, Rb1, Rc, Rb2, and Rd and their total in leaf, stem, and root of authentic wild-grown material. The totals in roots varied from 1 to 16%, with the greatest number of individual samples having 4-5% total ginsenosides. The lack of ginsenoside Rf in roots of authentic wild populations confirmed its status as a phytochemical marker differentiating American and Asian ginseng. Ten geographically isolated wild populations were collected, and several showed significant variation in the levels of major ginsenosides. There was no statistical difference in mean ginsenoside content between wild and cultivated P. quinquefolius roots at 4 years of age, suggesting there is no phytochemical justification for wild crafting. Baseline data on total ginsenoside levels for authentic wild P. quinquefolius reported here provide reference levels for quality assurance programs.     

Determination of ginsenosides Rb1, Rc, and Re in different dosage forms of ginseng by negative ion electrospray liquid chromatography-mass spectrometry

A method based on high-performance liquid chromatography (HPLC) and negative ion electrospray mass spectrometry (LC-MS) has been used to determine ginsenosides Rb1, Rc, and Re in six different samples of ginseng. These included a liquid extract, capsules, tea bags, and an instant tea. It was found that four of the six samples had detectable levels of at least one of the ginsenosides. The liquid extract, capsules, instant tea, and tea bags labeled ginseng had ginsenosides. The labels on the two samples that did not have ginsenosides indicated that they were a mixture of green tea, licorice, and ginseng. Also, 13C NMR was used to identify the types of complex carbohydrates present in the samples. One of the samples of tea bags had none of the ginsenosides, but did have complex carbohydrates found in most of the other samples. The instant tea had all three ginsenosides, but had no complex carbohydrates, only sucrose. The amounts of ginsenosides found in standard doses from six different sources of ginseng varied considerably. It was found that steeping a tea bag for a longer time than that recommended on the label produced a larger recovery of ginsenosides and that reusing a tea bag produced even higher recoveries.     

Simplified extraction of ginsenosides from American ginseng (Panax quinquefolius L.) for high-performance liquid chromatography-ultraviolet analysis

Four methods were tested for extraction and recovery of six major ginsenosides (Rb1, Rb2, Rc, Rd, Re, and Rg1) found in roots of American ginseng (Panax quinquefolius): method A, sonication in 100% methanol (MeOH) at room temperature (rt); method B, sonication in 70% aqueous MeOH at rt; method C, water extraction (90 degrees C) with gentle agitation; and method D, refluxing (60 degrees C) in 100% MeOH. After 0.5-1 h, the samples were filtered and analyzed by high-performance liquid chromatography (HPLC)-UV. A second extraction by methods C and D was done, but 85-90% of ginsenosides were obtained during the first extraction. Lyophilization of extracts did not influence ginsenoside recovery. Method D resulted in the highest significant recoveries of all ginsenosides, except Rg1. Method C was the next most effective method, while method A resulted in the lowest ginsenoside recoveries. Method B led to similar recoveries as method C. All methods used one filtration step, omitted time-consuming cleanup, but maintained clear peak resolution by HPLC, and can be used for quantitative screening of ginsenosides from roots and commercial ginseng preparations.     

Influence of zinc on the growth,distribution of elements,and metabolism of one‐year old American ginseng plants

Using a water culture technique, 0.05 ppm zinc (Zn) was found to be the critical deficiency concentraction for one‐year American ginseng (Panax quinquefolium L) plants, 0.3 ppm was optimum, 0.5 ppm the critical Zn‐toxicity concentration, and 10 ppm the concentration when severe toxicity occurs. Therefore, the optimum Zn concentration for the growth of American ginseng plants was between 0.1 ppm ‐ 0.3 ppm. Zinc deficiency symptoms of one‐year old American ginseng plants were indicated by the inhabition of root growth, with little fibrous root development, and smaller leaves compared to normal leaves. The symptoms of toxicity were also indicated by the inhibition of root growth, and when seedlings were suffering from an acute toxicity, no fibrous roots appeared, and eventually the roots yellowed and leaves grew slowly or even entirely ceased to grow, the final result being very small leaves which are also chlorotic. Zinc maintained within the 0.1 ppm to 0.3 ppm sufficiency range promoted the synthesis and accumulation of ginsenosides by American ginseng plants, and both low and high Zn concentrations restrained the synthesis and accumulation of ginsenosides. Both Zn deficiency and the optimum Zn concentration (0.3ppm) are beneficial to the accumulation of amino acids in the roots of American ginseng plants. Close to the optimum Zn concentration, the ratios of P/Zn and Fe/Zn in the shoot of American ginseng plants were maintained at 77 and 9.4, respectively.     

Effects of population, age, and cultivation methods on ginsenoside content of wild American ginseng (Panax quinquefolium)

Genotype and environmental effects on ginsenoside content among eight wild populations of American ginseng (Panax quinquefolium) were investigated. Root concentrations of six ginsenosides were determined at the time of collection of plants from the wild (T0) and 2 years (T2) after transplanting roots from each of the eight populations to each of two different forest garden locations. Both location and population had significant effects on root and shoot growth. Overall, ginsenoside Rb1 was most abundant, followed by Rg1 and Re. Concentrations of Rg1 and Re were inversely related among and within populations. The relative ranking of populations differed depending upon the particular ginsenoside and sampling time. The relative importance of genotype and environment was not the same for all ginsenosides. Ginsenoside Re was influenced by population but not location, whereas Rb1, Rc, and Rb2 were influenced only by location (environment), while Rg1 and Rd were influenced by both. Ginsenoside levels were consistently lower, but growth was consistently higher at the more intensively managed garden location.     

Study on the nitric oxide scavenging effects of ginseng and its compounds

In this study, an in vitro nitric oxide (*NO)-generating system was used to investigate the *NO-scavenging effects of methanolic extracts of white ginseng (Panax ginseng C.A. Meyer), red ginseng, and sun ginseng and several ginsenosides and phenolic compounds. Sun ginseng extract showed the strongest activity among the three ginseng extracts. None of the ginsenosides used in this experiment showed *NO-scavenging activity, but the phenolic compounds, such as p-coumaric and vanillic acids, and maltol inhibited *NO production in a concentration-dependent manner. Moreover, maltol levels markedly increased by heat processing. Therefore, the enhanced *NO-scavenging activity of ginseng by heat processing was closely related to phenolic acids and the increased content of maltol.     

Protopanaxatriol-type ginsenosides from the root of Panax ginseng

Ginseng, the root of Panax ginseng C. A. Meyer (Araliaceae), is one of the most important traditional medicines and functional foods. A detailed phytochemical investigation on the roots of P. ginseng led to the isolation of 6 new natural protopanaxatriol (PPT)-type ginsenosides, ginsenosides Re(1)-Re(6) (compounds 1-6), along with 10 known PPT-type ginsenosides. Their structures were elucidated on the basis of chemical and spectroscopic analyses, including high-resolution mass spectrometry (HRMS) and 1D and 2D nuclear magnetic resonance (NMR). The unusual α-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl sugar chain, as found in compounds 1 and 2, is reported in the genus Panax for the first time.     

内生真菌GG22诱导子对地黄毛状根生长和次生代谢的影响

为探索不同浓度内生真菌GG22诱导子与地黄毛状根共培养不同时间后,对毛状根的生长及次生代谢产物合成的影响,本试验通过制备地黄内生真菌GG22诱导子溶液,加入已培养23 d的毛状根液体培养基中,使含诱导子的多糖浓度分别为0(对照)、10、30、50和100 mg·L^-1,共培养0、3、5、8、11 d后,分别测定地黄毛状根的鲜重、干重、总环烯醚萜苷及毛蕊花糖苷的含量。结果表明,加入诱导子溶液的试验组毛状根的鲜重、干重、总环烯醚萜苷及毛蕊花糖苷的含量比对照组高。且通过变异系数权重法分析综合得分,得出最适宜诱导条件为诱导子浓度50 mg·L^-1,共培养11 d。由此推测地黄内生真菌GG22诱导子在一定诱导浓度和培养天数下能够促进地黄毛状根的生长,并提高其次生代谢产物的含量。本试验结果为进一步研究地黄内生真菌GG22诱导子促进地黄次生代谢产物积累的分子机制提供了基础。