Two new saponins from the aerial part of Astragalus membranaceus var. mongholicus

Two new saponins named mongholicoside A （1） and mongholicoside B （2） were isolated from the aerial part of Astragalus membranaceus var mongholicus. Their structures were determined by 1D and 2D NMR, ESI-MS techniques and chemical methods.

Study on Tissue Culture and Radiation Mutation of Astragalus membranaceus Bge. var. mongholicus(Bge.) Hsiao

[Objective] The research aimed to study the tissue culture technology and callus induction by radiation mutation of A. membranaceus Bge. [ Method ] With the different parts of Astragalus membranaceus Bge. var. mongholicus （ Bge. ） Hsiao aseptic seedling as explants （ leaves, cotyledons, hypocotyls） induced callus, and cotyledon and hypocotyls taken by the method of radiation mutation were studied. [ Result]The results showed that the three explants had relatively high callus induced rate in the medium which respectively made up of MS ＋6-BA 2.0 mg/L ＋ NAA2.0 mg/L, LS ＋6-BA2.0 mg/L ＋NAA0.1 mg/L, MS ＋ 6-BA2.0 rng/L ＋ NAA2.0 rag/L; the optimum mutation time of hypocotyls and cotyledons was 15 minutes; the growth of the callus induced from hypocotyls would be better as the mutation time increased, but when it reached a certain time the growth would be weaken, the induction rate also would be reduced. [ Conclusion] This study will provide the scientific reference in tissue culture and mutation breeding of A. membranaceus Bge.

Comparative analysis of twenty-five compounds in different parts of Astragalus membranaceus var.mongholicus and Astragalus membranaceus by UPLC-MS/MS

As a traditional Chinese medicine,the root of Astragalus membranaceus var.mongholicus(AMM) or A.membranaceus(AM) has been widely used in China and other Asian countries for thousands of years.Till now,the flavonoids,phenolic acids and saponins are considered as the main active components contributing to their therapeutic effect in these plants.In order to clarify the distribution and contents of these compounds in different organs of these plants,a rapid and sensitive analytical method for simultaneous determination of 25 active compounds including seven types(i.e.dihydroflavones,isoflavane,isoflavones,flavones.pterocarpans,phenolic acid and saponins) within 10 min was established using ultra-pressure liquid chromatography coupled with tandem mass spectrometry(UPLC-MS/MS).Then,the established method was fully validated and successfully applied to the determination of the contents of these analytes in different parts(root,rhizome,stem,leaf and flower) of AMM and AM.The results indicated that the contents of the same type of compounds in two different species plants were significantly different.Moreover,the obvious differences were also found for the distribution and contents of different type of compounds in five organs of the same species.The present study could provide necessary information for the rational development and utilization of AMM and AM resource.

Analysis of the distribution of Astragalus membranaceus var.mongholicus in Inner Mongolia under climate change using the GEE platform

Objective:Astragali Radix,commonly used in traditional Chinese medicine,is subjected to an analysis of the habitat distribution of Astragalus membranaceus var.mongholicus in the Inner Mongolia Autonomous Region of China under climate change.This analysis is crucial for the conservation ofmedicinal plants,the assessment of their adaptability to climate change,and the promotion of sustainable development in the ecological environment.Methods:This study explores the currently suitable habitats for A.membranaceus var.mongholicus using a random forest species distribution model on the Google earth engine platform.Based on future climate data,the study predicts distribution dynamics for 4 time periods(2021–2040,2041–2060,2061–2080,and 2081–2100)under three climate change scenarios(SSP126,SSP245,and SSP585).Results:The results indicate that under current climatic conditions,the suitable habitat distribution of A.membranaceus var.mongholicus is mainly concentrated in the southwestern region of Inner Mongolia.Under future climate change scenarios,there is a varying degree of contraction in suitable habitats,with notable reductions.Particularly,under the SSP126 scenario(2081–2100),there is a significant decrease of 22.77%in suitable habitat.The SSP245 scenario(2041–2060)exhibits the most pronounced habitat loss,with a reduction of 36.38%.Additionally,with climate change,the overall suitable range of A.membranaceus var.mongholicus is expected to shift toward higher latitudes and contract.Conclusion:This study can be used to improve conservation efforts for wild A.membranaceus var.mongholicus.It also provides a scientific basis for the cultivation of this medicinal plant in the context of climate change.This,in turn,can encourage more industries to achieve both sustainable development and economic benefits,providing valuable insights for building a low-carbon economy.

Content Determination of Astragaloside Ⅳ in Astragalus from Three Different Regions by HPLC

[Objectives] The content of astragaloside in Astragalus membranaceus(Fisch.) Bge.var.mongholicus(Bge.) Hisao from three different regions was determined.[Methods] Referring to the method recorded in the Chinese Pharmacopoeia(2015 edition),the content of astragaloside IV in A.membranaceus was determined by HPLC.[Results] There were great differences in the astragaloside IV content of A.membranaceus among different regions.The content of astragaloside IV in A.membranaceus cultivated in Inner Mongolia was highest(0.155%),followed by that(0.143%) in A.membranaceus cultivated in Gansu,and the content of astragaloside IV in A.membranaceus cultivated in Shanxi was lowest(0.080%).The contents of astragaloside IV in A.membranaceus from different regions were all in line with the standard(not less than 0.040%) of Chinese Pharmacopoeia(2015 edition).[Conclusions]The content of astragaloside IV in A.membranaceus cultivated in three different regions met the medicinal standards.

移栽期对蒙古黄芪(Astragalus membranaceus var.mongholicus)药材产量和质量的影响

采用初冬和春季移栽对比试验,定期测定土壤含水量、生长发育指标、药材产量和质量指标,旨在探寻地道产区蒙古黄芪(Astragalus membranaceus var.mongholicus)最佳移栽季节。结果表明:与春栽田比较,冬栽田20cm土层内土壤含水量在苗栽返青期显著增高,而40cm以下土层土壤含水量在生长后期显著降低;苗栽返青和开花分别提早10d和13d,返青出苗势、出苗率和出苗指数分别提高32.6%、4.4%和0.2497,返青株成活率提高5.0%,有效延长了黄芪生长天数,为药材根的生长和有效成分积累奠定了良好的营养生长基础。初冬栽和春栽田成药根数分别为201 087根·hm-2和181 250根·hm-2,一等品出成率分别为24.02%和19.69%,药材鲜产量分别为5 400kg·hm-2和5 057kg·hm-2;黄芪甲苷含量分别为0.121%和0.098%,毛蕊异黄酮葡萄糖苷含量分别为0.080%和0.052%,水溶性浸出物含量分别为27.62%和25.54%。初冬移栽黄芪成效更为显著,建议规范化生产中合理调整移栽季节,提倡初冬移栽。

蒙古黄芪育苗基质的筛选及应用潜力初步评价

为筛选出适宜培育高质量幼苗的育苗基质,促进移栽蒙古黄芪(Astragalus membranaceus var.mongholicus)的生产,以酒糟为主料,牛粪和沼渣为辅料,按不同体积比配制了6种有机复合基质,并对其理化性质,以及碱解氮、速效磷、速效钾的含量进行了测定。通过PLS-DA分析辅料对复合基质性质的影响,采用模糊评价法评估各复合基质肥效,进而筛选出基质F2(酒糟∶牛粪=60∶20)和Z2(酒糟∶沼渣=60∶20),将二者分别与纯土按体积比3∶7混合制成育苗基质后,通过发芽和幼苗生长试验评价应用潜力。结果表明,各有机复合基质理化性质基本符合常规基质要求,分别以牛粪和沼渣为辅料的两组复合基质,F2和Z2的肥效评价系数较高。混合育苗基质“F2+纯土”组的蒙古黄芪种子发芽率最高,且发芽势和发芽指数显著高于“Z2+纯土”组和对照组;同时,该组幼苗的整株长、株高和鲜重等指标也显著优于其他两组。由此可见,有机复合基质F2性能良好,与纯土混合后制成的育苗基质可用于蒙古黄芪优质幼苗的培育,具有较好的应用潜力。

干旱胁迫对蒙古黄芪生长及根部次生代谢物含量的影响

以山西道地药材黄芪一年生幼苗为试验材料,设置常规水分条件(CK)、轻度干旱胁迫(A1)、中度干旱胁迫(A2)、重度干旱胁迫(A3)4个不同处理,研究土壤干旱胁迫对黄芪生长及生理的影响。结果表明:黄芪茎叶快速生长集中在出苗后80—120d,以后生长减缓;当茎叶枯萎时,根中生物量短期快速积累。与常规水分条件相比,干旱胁迫处理显著降低了黄芪苗高及茎叶生物量,但对抗氧化能力、根系生长及次生代谢物积累产生了不同的影响。轻度干旱胁迫下SOD、POD、CAT 3种抗氧化酶活性升高,丙二醛(MDA)含量和细胞膜透性降低,同时根长与根生物量增加、多糖与皂苷两种次生代谢物积累增多,黄芪药材的质量得到显著提高(P<0.05);胁迫上升到中度、重度时,SOD酶活性逐渐降低,重度胁迫下低于对照,而POD及CAT酶活性、MDA含量、细胞膜透性均随胁迫增强而升高,相反,根长、根生物量、多糖与皂苷含量降低,导致黄芪药材的质量显著降低(P<0.01)。综上表明,干旱胁迫下,SOD酶表现较为敏感,可能在清除活性氧中起主要作用;轻度水分胁迫能有效启动黄芪体内抗氧化酶系统和次生代谢,它们相互协作共同对抗胁迫对细胞产生的伤害,通过降低地上部分的生长,将营养物质优先运往根部,促进根产量及药材质量的提高。这一结论,可在黄芪多糖和皂苷次生代谢物定向培育的水分管理中加以利用。

蒙古黄芪中黄酮类成分的研究

目的研究蒙古黄芪的化学成分,为该中药的开发利用和质量评价提供依据。方法利用多种色谱方法分离纯化,通过理化常数测定和波谱分析鉴定其化学结构。结果从蒙古黄芪中分离鉴定了9个黄酮类化合物,分别为芒柄花素(formononetin,Ⅰ)、(3R)-8,2′-二羟基-7,4′-二甲氧基异黄烷[(3R)-8,2′-dihydroxy-7,4′-dimethoxy-isoflavane,Ⅱ]、毛蕊异黄酮(calycosin,Ⅲ)、(6aR,11aR)9,10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷[(6aR,11aR)9,10-dimethoxypterocarpan-3-O-β-D-glucoside,Ⅳ]、7,2′-二羟基-3′,4′-二甲氧基异黄烷-7-O-β-D-葡萄糖苷(7,2′-di-hydroxy-3′,4′-dimethoxy-isoflavane-7-O-β-D-glucoside,)、芒柄花素-7-O-β-D-葡萄糖苷(formononetin-7-O-β-D-glucoside,Ⅴ)、毛蕊异黄酮-7-O-β-D-葡萄糖苷(calycosin-7-O-β-D-glucoside,Ⅵ)、红车轴草异黄酮-7-O-β-D-葡萄糖苷(pratensein-7-O-β-D-glucoside,Ⅶ)和染料木苷(genistin,(Ⅸ))。结论化合物为首次从黄芪属植物中分得,化合物为首次从该种植物中获得,化合物～具有促进细胞增殖的活性。-

干旱胁迫对黄芪植株生长中黄酮类成分积累的影响

以3年生蒙古黄芪植株为材料,采用盆栽法模拟干旱处理,研究水分胁迫对黄芪生理状态以及其根、茎、叶中毛蕊异黄酮、毛蕊异黄酮葡萄糖苷、芒柄花素和芒柄花苷4种黄酮类化合物含量的影响。结果显示:(1)毛蕊异黄酮和毛蕊异黄酮葡萄糖苷在黄芪根、茎、叶均有分布,不同器官的毛蕊异黄酮葡萄糖苷含量表现为根>叶>茎,而毛蕊异黄酮含量则是叶>茎>根。(2)干旱胁迫下,黄芪根中毛蕊异黄酮葡萄糖苷和芒柄花苷在胁迫第8~12天有一定的积累;叶中的毛蕊异黄酮则在干旱第4~8天有明显的积累,而毛蕊异黄酮葡萄糖苷则没有明显变化。研究发现,4种黄酮类化合物在黄芪各器官中分布和含量不尽相同,同一器官中不同黄酮类化合物对干旱胁迫的响应规律也存在差异;适度的干旱胁迫能够促进黄芪毛蕊异黄酮葡萄糖苷的积累,但过度干旱胁迫则不利于其积累。

砷胁迫对药用植物黄芪中抗氧化酶活性的影响

[目的]探讨砷胁迫对药用植物黄芪中抗氧化酶活性的影响,以期为揭示砷毒害植物机理提供理论依据。[方法]以内蒙药用植物黄芪为材料,分别采用高锰酸钾滴定法、愈创木酚法、邻苯三酚自氧化法以及硫代巴比妥酸法研究不同浓度砷胁迫下对药用植物黄芪中过氧化氢酶(CAT)、过氧化物酶(POD)、超氧化物歧化酶(SOD)的活性影响以和丙二醛(MDA)的浓度变化。[结果]随着砷浓度升高,黄芪叶片中这3种酶的活性能够维持,特别是POD活性显著增加。从时间动态看,随处理时间的延长,黄芪叶片中的CAT活性先升高再降低,POD活性增加,SOD活性变化不显著;MDA含量先升后降,第8天达到最大值。[结论]该研究可为中药材重金属含量控制和药理活性鉴定提供理论依据。

播种量和方式对甘肃渭源蒙古黄芪育苗质量和产量的影响

以陇芪2号为试验材料,采用不同播种量(90,150,210kg/hm^2)和播种方式(条播,撒播,覆膜穴播)两因素育苗,育苗结束后秋季测定种苗质量指标和产量,旨在揭示不同播种量和播种方式互作对蒙古黄芪种苗质量和产量的影响,为其规范化栽培提供依据。结果表明,不同播种量和播种方式交互条件下育苗,蒙古黄芪种苗个体质量和群体质量均存在显著差异。在90kg/hm^2最低播种量基础上,随着播种量的增加,种苗单根鲜重、根长和根粗等个体质量指标均呈降低趋势,种苗产量先极显著提高后下降,产苗数持续增加,优质种苗率显著下降。在同一播量条件下覆膜穴播方式育成种苗个体质量最优,优质种苗率均比条播和撒播高,并随播种量递增,苗产量较撒播和条播呈先增加后下降,而产苗数呈下降趋势。播种量150kg/hm^2条件下采用覆膜穴播育成单位面积优质种苗数最多。说明在保证种苗个体质量和单位面积产苗数的前提下,渭源蒙古黄芪育苗采用覆膜穴播方式,种子播种量为150kg/hm^2最佳。

高速逆流色谱法分离纯化黄芪中的芒柄花素和毛蕊异黄酮

采用高速逆流色谱法(HSCCC),以正己烷氯仿甲醇水组成二相系统作为固定相与流动相,对黄芪的乙酸乙酯粗提物进行了分离纯化。结果发现:以正己烷氯仿甲醇水(体积比为1.5∶3∶3∶2)组成的系统可以从黄芪的乙酸乙酯粗提物中分离出毛蕊异黄酮,纯度可达95%以上,并可以初步纯化芒柄花素;接着用正己烷氯仿甲醇水(体积比为4∶4∶5∶4)组成的系统进一步纯化芒柄花素,其纯度达95%以上。利用该方法,可以对中药黄芪中的异黄酮进行快速的分离和纯化。

黄芪多倍体的诱导与鉴定

本试验采用改良琼脂涂抹法(0.2％的秋水仙碱与0.1％的琼脂混合成半固体)处理黄芪Astragalusmembranaceus var.mongholicus幼苗顶芽,得到的诱导株,经生物学性状鉴定和染色体鉴定为四倍体。

黄芪皂苷纯化工艺研究

目的 :研究黄芪皂苷类成分的纯化方法。方法 :应用色谱指纹谱技术 ,研究比较正丁醇萃取法、大孔吸附树脂法纯化黄芪总皂苷。结果 :AB 8树脂吸附纯化与正丁醇萃取纯化的黄芪总皂苷得率相近 ,但指纹图谱有一定差别 ,而其它型号的树脂吸附纯化的总皂苷得率偏低。结论 :纯化黄芪总皂苷 ,AB 8树脂优于其它型号树脂。

蒙古黄芪对N、P、K的肥效响应

采用三因素二次D-饱和最优设计,研究N、P、K及其配比对沙地1a生蒙古黄芪的株高、根长、根直径、根干质量等生物学特性及皂苷、黄酮、多糖次生代谢物和Fe、Mn、Cu、Zn微量元素质量分数的影响,利用主成分分析其最佳施肥用量。结果表明:氮肥有助于黄芪株高的增高,但施肥量过高黄芪株高反而降低,以氮钾配施处理(N2K2)的株高最高,较对照增加51.93%;磷肥和钾肥能促进根的生长发育,其中氮、磷、钾施肥量分别为79.41、150和225kg/hm^2处理(N1P3K3)的效果最佳,其根直径与根干质量分别较对照增加58.42%和43.87%。施肥能显著提高蒙古黄芪皂苷和多糖的质量分数,以氮、磷、钾施肥量分别为225、150和79.41kg/hm^2处理(N3P3K1)的效果最好,其皂苷和多糖的质量分数分别较对照增加118.52%和13.14%;单施氮肥(N3)和单施磷肥处理(P3)均不利于黄酮的积累,其黄酮质量分数分别较对照降低7.56%和6.46%。对于微量元素,施肥能促进黄芪Mn的积累,对Cu、Zn影响不大,单施钾肥处理(K3)使得黄芪Fe元素质量分数降低。此外,结合蒙古黄芪的次生代谢物及微量元素的主成分分析,最终得出该地区蒙古黄芪的最佳施肥用量为氮(N)、磷(P2O5)、钾(K)分别为79.41、150和225kg/hm^2。

Analysis of principal isoflavone glycosides and aglycones in Radix astragali

Two major isoflavone glycosides [calycosin 7-O-β-D-glucopyranoside （1） and ononin （2）] and their aglycones [calycosin （3） and formononetin （4）] were simultaneously quantified with HPLC/DAD method. Two unknown compounds were identified as calycosin 7-O-β-D-glucopyranoside-6＇＂-O-malonate （U1） and formononetin 7-O-β-D-glucopymnoside-6＇＂-O-malonate （U2）, respectively, with LC/MS^n. Raw Radix astragli were shown to have higher contents of isoflavone glycosides （1, 2）, but lower contents of aglycones （3, 4） than the processed herbal materials. After being moistened with water and stored up for 24 h at 35 ℃, the glycosides and their m_alonates were almost completely transformed to their corresponding aglycones. The different contents of the isoflavone glycosides and their aglycones in raw and processed Radix astragali materials might be due to enzymolysis of the glycosides during processing.

蒙古黄芪植株水浸液的自毒作用研究

目的:通过对蒙古黄芪植株各部位提取液的生物学试验,探讨蒙古黄芪是否存在自毒作用,以及自毒作用与连作障碍的关系。方法:设置蒸馏水(CK)、蒙古黄芪根、茎和叶水浸提液各12.5、25、50和100 mg/mL 4个浓度,共计13个处理,用于黄芪自毒作用的生物学实验。结果:当蒙古黄芪根、茎和叶水浸提液的浓度达到25 mg/mL时,对自身种子萌发和幼苗生长有抑制作用,且随水浸提液浓度的升高,这种抑制作用增强;就综合化感效应而言,茎水浸提液的抑制作用>叶水浸提液>根水浸提液。发芽指数和幼苗活力指数对水浸提液比其他指标更为敏感。结论:蒙古黄芪植株水浸提液对自身种子萌发和幼苗生长具有明显的抑制作用,且抑制作用具有一定的浓度依赖性,蒙古黄芪自毒作用是造成连作障碍的原因之一。

火焰原子吸收法测定不同产地栽培黄芪中13种金属元素含量

目的:应用火焰原子吸收法测定不同产地栽培黄芪中的金属元素 Na、K、Mg、Ca、Cu、Zn、Mn、Fe、Co、Ni、Cd、Cr、Pb 含量。方法:采用先灰化、再用体积比为4:1的硝酸-高氯酸常压微沸条件下消解黄芪样品,研究了测定不同元素的仪器最佳工作条件,并作了方法的准确性和精密度考察。结果:河东、河西栽培黄芪中 Na、K、Mg、Ca、Mn、Fe、Cu、Zn 含量分别为1571.2,3962.0,2057.8,2968.0,10.4,406.7,8.2,12.2 μg·g^(-1)和227.2,11274.8,1047.1,983.5,10.5,362.3,11.3,19.9 μg·g^(-1)。Co、Ni、Cd、Cr、Ph 未检出。方法的加样回收率为97.16%～102.7%,精密度 RSD(n=9)为0.5%～2.6%。结论:测定方法简单易行,方便快捷,结果令人满意。

中药材产地适宜性分析地理信息系统的开发及蒙古黄芪产地适宜性研究

依托地理信息系统(GIS)平台,以气温、相对湿度、降水量、日照时数、极端最低温度、极端最高温度的1km^2的栅格气候数据库以及1：4000000的土壤数据库,并以1：1000000的基础地理信息数据为基础,利用聚类分析和空间分析技术,建立了《中药材产地适宜性分析地理信息系统》(TCMGIS-Ⅰ)。以内蒙古武川和山西浑源为道地基点县,运用《中药材产地适宜性分析地理信息系统》(TCMGIS-Ⅰ),分析了蒙古黄芪的全国适宜产地。结果表明按蒙古黄芪药材生长所需要的气候、土壤条件分析,蒙古黄芪主要适宜产区集中分布在山西北部和内蒙古的南部,黄河以南的华东地区基本没有适宜区。系统分析结果对合理发展蒙古黄芪生产、认识黄芪道地产地形成具有重要指导意义。