Chemical Constituents of Glycyrrhiza eurycarpa P. C. Li (1I)

Huang Gancao (Glycyrrhiza eurycarpa P. C. Li) is a new species discovered recently. Its chemical constituents have not been reported before. This paper reports the isolation of ten components (Ⅰ—Ⅹ) from its roots and rhizomes collected in Jinta County, Gansu Province. On the basis of physico-chemlcal constants and spectroscopic (IR, UV, 1HNMR, 13CNMR, MS, and CD) analysis, they were identified as ononin(Ⅰ), liquiritigenin 4 -O-β-D-apio-D-franosyl-(1 →2)-β-D-glucopyranoside(II), isoliquiritigenin-4-O-β-D-apio-D-franosyl-(1→ 2) β-D-glucopyranoside (Ⅲ), choerospondin (Ⅳ), violanthin (Ⅴ), isoviolan thin (Ⅵ), vicenin-2(Ⅶ), uralsaponin A(Ⅷ), daucosterol (Ⅸ), and phloretic acid (Ⅹ), respectively All of them were isolated from this plant for the first time. Choerospondin, violanthin, daucosteroi and phloretic acid were not found in this genus before.     

新疆甘草属(Glycyrrhiza) 的生态分布及其经济利用

 新疆分布的甘草属植物共五种：甘草(Clycyrrhiza uralensis)、光果甘草(G．glabra)、粗毛甘草(G. aspera)、胀果甘草(G．inflata)、科氏甘草(G．korshinskyi)。出现于新疆88％的县市,各大河流沿岸、泉水溢出带及冲积平原。在区系上与中亚关系密切。空气干燥度、年平均温度及土壤盐碱含量对不同种甘草的分布都有影响。甘草虽然在南、北、东疆都有分布,但以北疆为主。甘草分布地区的大气干燥度从1.37—237.3,而以3—6最佳;年均温度为0.4—14℃,而以4—8℃为最适宜。甘草常与苦豆子(Sophora alopecuroides)、芨芨草(Achnatherum splendens)、芦苇(Phragmites communis)、大叶白麻(Poacynum hendersonii)等组成群落。光果甘草可与灰杨(Populus pruinosa)、柽柳(Tamarix chinensis)、假苇拂子茅(Calamagrostis Pseudophragmites)等组成群落。胀果甘草常与不同比例的胡杨(Populus diversifolia)、铃铛刺(Halimodendron halodendron)、柽柳、芦苇、大叶白麻、骆驼刺(Alhagi pseudalhagi)、花花柴(Karelinia caspica)等组成群落。粗毛甘草多为田间、果园及路旁荒地杂草。为了保持甘草资源的持续利用并充分发挥其效益,应建立合理的挖采制度,建立甘草生产基地,并开展甘草的综合利用研究。     

新疆胀果甘草总黄酮的细胞毒活性研究

为了探讨新疆胀果甘草总黄酮(general flavonoids,GFs)对宫颈癌细胞的细胞毒活性,自制备胀果甘草GFs,通过离子阱喷雾式质谱技术分析,建立GFs组分的指纹图谱;并且对SiHa宫颈癌细胞进行GFs药物干预,应用MTT法和流式细胞技术分别鉴定细胞活力和细胞凋亡率。获得了新疆胀果甘草GFs组分的特征性指纹图谱,提取物中GFs含量可达35.40%;发现在0～500μg/mL GFs浓度范围内,GFs药物干预引起细胞活力梯度性下降,其MTT法检出的最低值为12%;在0～1000μg/mL GFs浓度范围内,GFs诱导的细胞凋亡率呈现梯度性上升趋势,其流式细胞仪检出的最高值为78%。结果表明新疆胀果甘草GFs具有较强的细胞毒活性,能够抑制宫颈癌细胞生长与活力,并诱导细胞凋亡。     

Comparative metabolite profiling and fingerprinting of medicinal licorice roots using a multiplex approach of GC–MS,LC–MS and 1D NMR techniques

Glycyrrhiza glabra, commonly known as licorice, is a popular herbal supplement used for the treatment of chronic inflammatory conditions and possesses anticancer and antiviral activities. This species contains a plethora of phytochemicals including terpenoids, saponins, flavonoids, polyamines and polysaccharides. The full complement of bioactive compounds has yet to be elucidated, a step necessary in order to explain its medicinal use. There are over 30 species in the Glycyrrhiza genus world-wide, most of which have been little characterized in terms of phytochemical or pharmacological properties. Here, large scale multi-targeted metabolic profiling and fingerprinting techniques were utilized to help gain a broader insight into Glycyrrhiza species chemical composition. UV, MS and NMR spectra of extracted components were connected with NMR, MS, and multivariate analyses data from Glycyrrhiza glabra, Glycyrrhiza uralensis, Glycyrrhiza inflata and Glycyrrhiza echinata. Major peaks in ¹H NMR and MS spectra contributing to the discrimination among species were assigned as those of glycyrrhizin, 4-hydroxyphenyl acetic acid, and glycosidic conjugates of liquiritigenin/isoliquiritigenin. Primary metabolites profiling using GC–MS revealed the presence of cadaverine, an amino acid, exclusively found in G. inflata roots. Both LC–MS and NMR were found effective techniques in sample classification based on genetic and or geographical origin as revealed from derived PCA analysis.     

Production of glycyrrhizin in callus cultures of licorice

Licorice plants, Glycyrrhiza glabra, G. uralensis, and G. inflata, were investigated for callus induction using Murashige and Skoog (MS) medium combined with auxins and cytokinins. After 4 weeks of culture, 33-100% of leaf or stem explants formed calli. Maximum of shoot induction from callus cultures was achieved by G. inflata stem explants cultured on MS medium supplemented with 1 mg/l alpha-naphthaleneacetic acid (NAA) and 0.5 mg/l 6-benzyladenine (BA) (67%) which also gave maximum shoot formation per explant (two shoots per explant). These results indicated that all three Glycyrrhiza species regenerated shoots from callus cultures on MS medium combined with NAA and BA or only thidiazuron (TDZ; 0.1 and 0.5 mg/l). Glycyrrhizin contents of G. uralensis calli induced using MS medium in combination with NAA and BA [(27.60 +/- 8.47) microg/g DW] or TDZ alone [(36.52 +/- 2.45) microg/ g DW] were higher than those found in other combinations.     

甘肃悬钩子属系统分类及区系地理研究

甘肃省产悬钩子属植物３４种、８变种、隶属于３组１１亚组,其中５个种为甘肃分布新记录、个新种及１个新变种。本文根据实地调查和标本鉴定,编制了系统检索表,并对甘肃悬钩子属植物区系成分及区系成进行了初步探讨。     

甘草属种间杂交种叶绿体DNA父系遗传的发现及分析

通过对甘草属乌拉尔甘草(Glycyrrhiza uralensis)、光果甘草(G.glabra)、胀果甘草(G.inflata)及其人工杂交种组合G.uralensis♀×G.glabra♂、G.glabra♀×G.uralensis♂、G.uralensis♀×G.inflata♂、G.inflata♀×G.uralensis♂共68份材料的核基因ITS序列、叶绿体rbc L、mat K、trn H-psb A基因的序列分析,探讨了甘草属叶绿体DNA遗传方式。结果表明:(1)亲本种和人工杂交种ITS序列长度均为614 bp,其中34份人工杂交种ITS序列存在4处变异位点,且人工杂交种均检测出来自父本、母本ITS序列相同位点碱基的叠加,检测率为100%。(2)亲本种与人工杂交种的叶绿体基因rbc L、mat K、trn H-psb A序列长度相同,共有4处变异位点,人工杂交种在变异位点处的碱基与其相对应的父本碱基一致率高达97.1%。以上结果说明,该研究获得34份人工杂交种为100%杂交成功的F_1子代,核基因ITS序列可用于甘草属杂交种的遗传鉴定;甘草属叶绿体rbcL、mat K、trn H-psb A基因具有父系遗传特性,推测甘草属质体的遗传方式主要表现为父系遗传,这种质体遗传方式的发现为甘草属杂交种和遗传多样性研究提供了新的认识,也为杂交种的亲本鉴定提供分子依据。     

国产甘草属植物的RAPD分析及其分类学研究

应用RAPD技术,探讨甘草属(G ly cy rrh iza L.)13种1变种30个植物类群的遗传差异和几个争议种的分类地位。从60个随机引物中筛选出14个多态性好的引物进行RAPD实验,DNA片段的二态数据用U PGM A聚类法构建系统发育树。共扩增出250条带,多态性带204条,约占总数的81.7%。聚类结果显示RAPD分子标记构建的系统发育树与经典分类系统一致。甘草属植物具有丰富的遗传多样性,同种内不同居群间的遗传分化较大。黄甘草、胀果甘草、乌拉尔甘草三者亲缘关系较近,平卧甘草与粗毛甘草存在很大的遗传差异,作为独立种较合理。RAPD标记可为甘草属植物的系统分类研究提供分子生物学依据。     

5种甘草属植物花序和种子生产的位置效应及繁殖资源分配模式初步研究

以种植于新疆石河子的光果甘草( Glycyrrhiza glabra Linn.)、胀果甘草( G. inflata Batal.)、乌拉尔甘草( G. uralensis Fisch.)、黄甘草( G. eurycarpa P. C. Li)和蜜腺甘草( G. glabra var. glandulosa X. Y. Li)为研究对象,对植株不同部位的花序数量、花序正常发育率、每花序单花数量和果穗干质量,以及植株不同部位和花序不同部位的生物量投入比、座果率、结籽率、种子投影面积和种子千粒质量进行测定;在此基础上,对供试5种甘草属( Glycyrrhiza Linn.)植物的繁殖资源分配模式和种子生产策略进行分析。结果表明：同一植株内,光果甘草、乌拉尔甘草、黄甘草和蜜腺甘草的花序数量、花序正常发育率、每花序单花数量和果穗干质量从植株下部到上部总体上依次递减,而胀果甘草植株不同部位间这4项指标总体上无显著差异。同一植株内,胀果甘草植株中部的生物量投入比和座果率均较高,但其生物量投入比、座果率和结籽率在植株不同部位间均无显著差异;而供试另4种植物的生物量投入比、座果率和结籽率从植株下部到上部总体上依次递减。同一花序内,胀果甘草花序中部的生物量投入比明显高于花序上部和下部,座果率从花序下部到上部依次递减,结籽率则在花序不同部位间无显著差异,而供试另4种植物的生物量投入比、座果率和结籽率从花序下部到上部总体上依次递减。供试5种植物的种子投影面积和种子千粒质量在植株不同部位间和花序不同部位间均无显著差异。综合研究结果显示：在资源竞争、结构效应和花粉限制的影响下,供试5种甘草属植物存在2种不同的资源分配模式和种子生产格局。其中,光果甘草、乌拉尔甘草、黄甘草和蜜腺甘草通过减少对晚发育的花或果实的资源投入来保障早发育的花或果实获得较多的资源,达到繁殖成功的目的;而胀果甘草则采取对花和果实随机败育的方式减小资源竞争的压力,这2种繁殖资源分配模式和种子生产策略对提高甘草属植物的繁殖成功率具有重要作用。     

新疆野生胀果甘草内生细菌多样性的非培养初步分析

摘要：【目的】了解新疆野生甘草内生细菌多样性,为开发新的微生物资源奠定基础。【方法】采用改进的CTAB (十六烷基三甲基溴化铵)法提取新疆野生胀果甘草根部总DNA,利用细菌16S rDNA 基因通用引物对甘草总DNA 进行16S rDNA 基因扩增,构建甘草内生细菌16S rDNA基因文库;挑选具有不同酶切图谱的克隆进行测序、比对并构建16S rDNA 基因系统发育树。【结果】构建的甘草内生细菌16S rDNA基因文库中, 150个克隆分属于32个不同的分类单元,Blast结果表明大部分克隆与已知细菌的16S rDNA基因序列相似性较高,分别归属于变形杆菌门(Proteobacteria)的alpha、gamma亚群,厚壁菌门(Firmicutes),放线菌门(Actinobacteria),拟杆菌门(Bacteroidetes)中的鞘脂菌属(Sphingobium),叶杆菌属(Phyllobacterium),生丝单胞菌属(Hyphomonas),土壤杆菌属(Agrobacterium)等14个属, 其中26%的克隆与已知细菌16S rDNA 基因相似性小于96%,可能代表新的分类单元.【结论】甘草内生细菌多样性丰富且存在尚未被认识的新物种。     

甘肃省忍冬属植物系统分类

根据文献资料,经过大量标本鉴定整理,确认甘肃省共有忍冬属植物34种、4亚种、6变种,其中包括1个新异名、2个新等级和1个新记录变种。     

四种前体对胀果甘草细胞悬浮培养生产甘草黄酮的调控效果评价

在胀果甘草细胞悬浮体系中加入苯丙氨酸、酪氨酸、肉桂酸和乙酸钠来研究前体对甘草细胞生产甘草黄酮的影响。结果显示,4种前体在合适的浓度下对细胞的生长没有明显的抑制作用,而且均能促进细胞内甘草黄酮的生物合成,但高浓度的肉桂酸对细胞的生长有一定的抑制作用。苯丙氨酸的最佳添加浓度为20 mg/L,酪氨酸、肉桂酸、乙酸钠的最佳添加浓度都是5 mg/L。此时,均可使培养体系的甘草黄酮产量高达100 mg/L以上,其中酪氨酸的添加使得产量高达对照的1.43倍。苯丙氨酸、肉桂酸和乙酸钠3种前体的添加时间均以第10天为宜,酪氨酸添加时间以第5天为最佳。而且,在添加苯丙氨酸和乙酸钠后的第3天收获细胞,此时细胞的生物量和甘草黄酮产量最大。此外,苯丙氨酸、乙酸钠的添加可以增加黄酮合成的关键酶之一——苯丙氨酸裂解酶的活性。酪氨酸对苯丙氨酸裂解酶影响不大,而肉桂酸的添加却导致其活性显著降低。     

Karyotypical Study of Chinese Species of the Genus Glycyrrhiza

Seven species of Glycyrrhiza in China are karyologically analyzed in this paper. The results are as follows: G. uralensis Fisch. 2n=16=6m+10sm; G. eurycarpa P.C. Li 2n=16=8m+6sm+2st; G. glabra L. 2n=16=14m+2sm; G. inflata Bat. 2n=16=6m+10 sm; G. aspera Pall. 2n=16=12m+4sm; G. yunnanensis Cheng f. et L. K. Tai ex P. C. Li 2n =16=12m+4sm; G. pallidiflora Maxim. 2n=16=12m+4sm. From this work and the previous reports, it is presumed that the basic chromosome number of Glycyrrhiza is x=8. Based on the characters of karyotype so far realed, the authors consider that the genus Glycyrrhiza is a relatively primitive group in Leguminosae and G. pallidiflora is the most primitive one among the species dealt with in this paper, and G. eurycar-pa is the most advanced one.     

不同耐盐性药用甘草幼苗根对Na~+的响应及其维管组织变化

以甘草属2种耐盐植物胀果甘草、乌拉尔甘草为材料,用不同浓度(50、100、150、200、250mmol·L-1)NaCl处理幼苗21d后,分析其生物量和根、茎、叶中的Na+、K+含量以及K+/Na+,计算根的离子选择吸收和运输系数,并应用光学显微镜观察比较二者的维管组织结构变化,以揭示2种药用甘草幼苗根对Na+的响应及其维管组织结构的变化特征,探讨甘草的耐盐机理。结果表明:(1)NaCl胁迫使2种甘草幼苗生物量均下降,在NaCl浓度为250mmol·L-1时,胀果甘草、乌拉尔甘草幼苗生物量分别是对照的53.34%、46.21%,胀果甘草耐盐性强于乌拉尔甘草。(2)随着NaCl浓度上升,2种甘草根积累的Na+显著增多,其中胀果甘草在所有盐处理下,根Na+含量均高于其它器官,说明其根对吸收的Na+具有显著截留效应;而乌拉尔甘草只在0~150mmol·L-1 NaCl范围内,根Na+含量显著高于叶片,当NaCl为200、250mmol·L-1时,叶片Na+含量显著高于根,说明乌拉尔甘草根对Na+的截留能力有限。(3)在相同盐处理下,胀果甘草离子选择吸收系数SAK,Na、离子运输系数STK,Na均大于乌拉尔甘草,胀果甘草根抑制Na+、促进K+向地上部运输的能力强于乌拉尔甘草。(4)乌拉尔甘草在NaCl为150、200mmol·L-1、胀果甘草在250mmol·L-1时,根结构对盐胁迫产生应激性响应,维管组织比例显著上升,有助于提高根向上的运输能力,减少盐害。研究表明,2种药用甘草根对Na+截留作用和向上运输时促K+抑Na+能力的差异,是导致其耐盐能力不同的主要原因,根对Na+的积累和截留作用的差异与根的结构响应相吻合,能较好地解释二者的耐盐性差异。     

甘草悬浮细胞的玻璃化法超低温保存研究

以甘草悬浮细胞为材料,进行了超低温保存技术的研究。高产黄酮的甘草悬浮细胞是一种极具价值但又难于保存的材料,为解决甘草细胞超低温保存中存在的关键问题,即如何减少渗透压的突然改变对细胞造成损伤,通过优化预培养时间、细胞年龄、预处理时间、脱水时间、洗涤液蔗糖浓度等影响因素,建立了一套适合于甘草悬浮细胞的保存方法。采用改良的2,3,5-氯化三苯基四氮唑(TTC)法检测,此法存活率可达82.9%。将保存后的细胞进行恢复培养,其恢复生长率可达80.4%。     

Methyl jasmonate elicitation enhances glycyrrhizin production in Glycyrrhiza inflata hairy roots cultures

Hairy roots were induced by infecting stems and leaves of Glycyrrhiza inflata with Agrobacterium rhizogenes ATCC 15834. The optimization of growth and glycyrrhizin accumulation of G. inflata hairy roots was studied. Sucrose (6%, w/v) was optimal for growth and glycyrrhizin accumulation in G. inflata hairy roots. Effects of elicitors like chitosan, methyl jasmonate, and yeast extract on glycyrrhizin production were studied. Methyl jasmonate (100 microM) was most efficient in enhancing glycyrrhizin production up to almost 109 microg/g dry weight on day 5 of elicitation. These results indicate that application of elicitors can enhance the capacity of G. inflata hairy roots to produce glycyrrhizin.     

3种甘草属植物的核型研究

用光学显微镜观察了豆科（Leguminosae）甘草属（Glycyrrhiza）3种荒漠植物的染色体,研究结果表明,3种植物的体细胞染色体数目2n=16,核型公式分别为：刺果甘草（G.pallidiflara）k(2n)=2x=16=4M+8m+4sm,光果甘草（G.glabra）k(2n)=2x=16=10M+6m,胀果某草（G.inflata）k(2n)=2x=16=6M+6m+4sm。     

新疆胀果甘草内生细菌枯草芽胞杆菌产甘草酸类代谢产物的初步研究

为筛选到可产生甘草酸的内生茵,从健康的胀果甘草(Glycyrrhiza inflata Bat.)不同组织中分离得到149株内生细菌,采用发酵培养的方法,以甘草酸单铵盐为标准品采用TLC法和HPLC法对这些细菌的代谢产物进行筛选,得到一株可能产甘草酸类似物的内生枯草芽胞杆菌Bacillussubtilis.     

Anti-Helicobacter pylori flavonoids from licorice extract

Licorice is the most used crude drug in Kampo medicines (traditional Chinese medicines modified in Japan). The extract of the medicinal plant is also used as the basis of anti-ulcer medicines for treatment of peptic ulcer. Among the chemical constituents of the plant, glabridin and glabrene (components of Glycyrrhiza glabra), licochalcone A (G. inflata), licoricidin and licoisoflavone B (G. uralensis) exhibited inhibitory activity against the growth of Helicobacter pylori in vitro. These flavonoids also showed anti-H. pylori activity against a clarithromycin (CLAR) and amoxicillin (AMOX)-resistant strain. We also investigated the methanol extract of G. uralensis. From the extract, three new isoflavonoids (3-arylcoumarin, pterocarpan, and isoflavan) with a pyran ring, gancaonols A[bond]C, were isolated together with 15 known flavonoids. Among these compounds, vestitol, licoricone, 1-methoxyphaseollidin and gancaonol C exhibited anti-H. pylori activity against the CLAR and AMOX-resistant strain as well as four CLAR (AMOX)-sensitive strains. Glycyrin, formononetin, isolicoflavonol, glyasperin D, 6,8-diprenylorobol, gancaonin I, dihydrolicoisoflavone A, and gancaonol B possessed weaker anti-H. pylori activity. These compounds may be useful chemopreventive agents for peptic ulcer or gastric cancer in H. pylori-infected individuals.