Terpenoids are widely distributed in plants and are often used as defense molecules in plant-microbe interactions. However, endophytic microorganisms usually establish a better symbiotic relationship with their hosts by secreting enzymes to avoid defense plant metabolites. This study evaluated the in vitro biotransformation activity of licorice endophytic fungi on glycyrrhizin and further explored the molecular regulation of their in vivo colonization on the licorice growth and metabolism. The results indicated that licorice endophytic fungi generally possessed the ability to bio-transform glycyrrhizin, with Z6 and Z15 exhibiting glycyrrhizin-induced β-glucuronidase activity. The Z6GH2 and Z15GH2 proteins were identified to hydrolyze glycyrrhizin in different ways by prokaryotic and eukaryotic experiments. In vivo re-infestation of licorice by Z6 and Z15 revealed significant promotion of glycyrrhizin biosynthesis and accumulation by regulating the expression levels of genes involved in glycolysis and glycyrrhizin biosynthesis pathway in licorice. These findings were further validated in J3, which has glycyrrhizin biotransformation properties. In summary, this study reveals the molecular mechanism by which endophytic fungi with glycyrrhizin β-glucuronidase activity promote glycyrrhizin biosynthesis and accumulation in licorice through feedback regulation of its metabolic flux. These finding highlight the importance of endophytic fungi in regulating the accumulation of active ingredients in medicinal plants.

The content of 15 total amino acids(TAAs) in Bambusae Concretio Silicea was determined by HPLC with phenyl-isothiocyanate(PITC) for pre-column derivatization. The results showed that the content of TAA was 0.61-12.25 mg·g~(-1), and aspartic acid(Asp), glutamic acid(Glu), proline(Pro), glycine(Gly), and valine(Val) were the top five amino acids in terms of the average content. The content of essential amino acids(EAAs), conditionally essential amino acids(CEAAs), non-essential amino acids(NEAAs), and medicinal amino acids(MAAs) was 0.24-4.75, 0.30-4.73, 0.40-7.50, and 0.36-6.51 mg·g~(-1), respectively. Among the delicious amino acids, sweet amino acids(SAA), bitter amino acids(BAA), fresh-taste amino acids(FAAs), and odourless amino acids(OAAs) had the content of 0.22-4.70, 0.19-4.03, 0.13-2.26, and 0.06-1.26 mg·g~(-1), respectively. The 21 batches of Bambusae Concretio Silicea samples presented the same composition but significant differences in the content of amino acids. Among the three producing areas, Guangdong was the area where the samples had the highest content of TAAs, EAAs, CEAAs, NEAAs, MAAs, and delicious amino acids. Furthermore, the ratio of amino acid(RAA), ratio coefficient of amino acid(RCAA), and score of ratio coefficient of amino acid(SRCAA) were calculated to evaluate the nutritional value of Bambusae Concretio Silicea. The results showed that the Bambusae Concretio Silicea samples from Guangdong had better nutritional value. The nutritional value evaluation based on the content of 15 amino acids was proposed to provide data support for the quality grading of Bambusae Concretio Silicea and lay a foundation for the development and utilization of the medicinal material resources.

The plant microbiome is one of the key determinants of healthy plant growth. However, the complexity of microbial diversity in plant microenvironments in different regions, especially the relationship between subsurface and aboveground microorganisms, is not fully understood. The present study investigated the diversity of soil microorganisms in different regions and the diversity of microorganisms within different ecological niches, and compared soil microorganisms and endophytic microorganisms. 16 S and ITS sequencing was used to sequence the soil and endophytes microbiome of honeysuckle. Alpha diversity analysis and principal component analysis (PCoA) were used to study the soil and endophyte microbial communities, and the function of endophyte bacteria and fungi was predicted based on the PICRUST2 process and FUNGuild. In total, there were 382 common bacterial genera and 139 common fungal genera in the soil of different producing areas of honeysuckle. There were 398 common bacterial genera and 157 common fungal genera in rhizosphere soil. More beneficial bacteria were enriched in rhizosphere soil. Endophytic bacteria were classified into 34 phyla and 770 genera. Endophytic fungi were classified into 11 phyla and 581 genera, among which there were significant differences in the dominant genera of roots, stems, leaves, and flowers, as well as in community diversity and richness. Endophytic fungal functions were mainly dominated by genes related to saprophytes, functional genes that could fight microorganisms were also found in KEGG secondary functional genes. More beneficial bacteria were enriched in rhizosphere soil of honeysuckle, and the microbial network of the rhizosphere is more complex than that of the soil. Among the tissues of honeysuckle, the flowers have the richest diversity of endophytes. The endogenous dominant core bacteria in each part of honeysuckle plant have a high degree of overlap with the dominant bacteria in soil. Functional prediction suggested that some dominant core bacteria have antibacterial effects, providing a reference for further exploring the strains with antibacterial function of honeysuckle. Understanding the interaction between honeysuckle and microorganisms lays a foundation for the study of growth promotion, quality improvement, and disease and pests control of honeysuckle from the perspective of microorganisms.

Drought adaptation of plants is closely related to resistance and tolerance to drought stress as well as the ability to recover after the elimination of the stress. Glycyrrhiza uralensis Fisch is a commonly applied herb whose growth and development are greatly affected by drought. Here, we provide the first comprehensive analysis of the transcriptomic, epigenetic, and metabolic responses of G. uralensis to drought stress and rewatering. The hyper-/hypomethylation of genes may lead to up-/downregulated gene expression, and epigenetic changes can be regarded as an important regulatory mechanism of G. uralensis under drought stress and rewatering. Moreover, integrated transcriptome and metabolome analysis revealed that genes and metabolites involved in pathways of antioxidation, osmoregulation, phenylpropanoid biosynthesis, and flavonoid biosynthesis may regulate the drought adaptation of G. uralensis. This work provides crucial insights into the drought adaptation of G. uralensis and offers epigenetic resources for cultivating G. uralensis with high drought adaptation.

目的:初步探究滇鸡血藤水煎液活血功效的生物评价方法并挖掘参与凝血酶抑制的核心成分.方法:采用体外凝血酶活性抑制试验,评价滇鸡血藤水煎液体外凝血酶抑制活性.然后利用超高效液相色谱-质谱(UPLC-MS)挖掘滇鸡血藤水煎液中的化学成分,并通过分子对接技术对这些化学成分的生物活性进行评估,筛选核心抗凝成分.结果:凝血酶抑制试验表明滇鸡血藤的标准煎剂(0.2 g·mL-1)具有高达78.40％的凝血酶抑制率,并从滇鸡血藤水煎液中推测鉴定了14个化合物,将其与凝血酶进行分子对接评价:戈米辛C(-7.9 kcal·mol-1)、戈米辛G(-7.6 kcal·mol-1)、内南五味子素C(-8.1 kcal·mol-1)、内南五味子素D(-9.4 kcal·mol-1)、阿里山五味子灵C(-9.4 kcal·mol-1)、五味子素C(-7.9 kcal·mol-1)、五味子素A(-7.3 kcal·mol-1)、异型南五味子素E(-8.2 kcal·mol-1)、五味子酯D(-8.6 kcal·mol-1)、红花五味子素(-7.4 kcal·mol-1)、内南五味子酯B(-7.6 kcal·mol-1)、内南五味子酯A(-8.1 kcal·mol-1)、乙酰基氧代南五味子烷(-8.5 kcal·mol-1)、内南五味子酯D(-9.2 kcal·mol-1).其中内南五味子素D和阿里山五味子灵C与凝血酶蛋白具有紧密结合的空间构型以及理想的结合能.结论:滇鸡血藤水煎液具有较好的凝血酶抑制活性,筛选出的2个具有凝血酶抑制活性的核心成分,可作为其质量标志物候选成分,为滇鸡血藤水煎液生物活性评价方法的建立和质控研究提供了参考依据.

Licorice is a product with the same origin as medicine and food and is widely used in traditional Chinese medicine and the food, cosmetics and tobacco industries. With the increasing demand for licorice in the herbal market, there is also increasing concern about complex varieties and quality deterioration. Considering the importance of the quality and safety of commercial licorice products, we used DNA barcoding, HPLC and colorimetry to distinguish and evaluate commercial licorice products. Here, 52 samples could be accurately identified into three kinds of licorice, Glycyrrhiza uralensis Fisch. (Gu), Glycyrrhiza inflata Bat. (Gi) and Glycyrrhiza glabra L. (Gg), by internal transcribed spacer (ITS) and trnH-psbA sequences. Among them, the S27 sample was identified as Gi but had been incorrectly labeled as Gg. Based on HPLC combined with stoichiometric analysis, the contents of liquiritin, glycyrrhizic acid and isoliquiritin contents were significantly different between Gu and Gg, and Gi and Gg showed significant differences only in isoliquiritigenin content. The chemical components and color difference parameters showed that isoliquiritigenin had a significant correlation with the color parameter b*. The above results show that DNA barcoding can quickly and accurately distinguish species, but it cannot provide data on metabolites or other quality information beyond the plant raw materials. Techniques that combine of DNA barcoding, HPLC and colorimetry can more easily distinguish and evaluate the types and quality of commercial licorice products. This study provides good data support for the safety and quality of licorice in the Chinese market.

By studying various ancient texts such as herbal classics and medical literature from different eras,it was found that there were discrepancies in the records about Bambusae Concretio Silicea(Tian Zhu Huang).In order to establish an accurate foundation,this research was based on ancient herbal literature and combined with plant morphology and investigative studies to examine its earliest mentions in ancient texts,nomenclature,medicinal properties,indications,and quality assessment standards.In the early records,Bambusae Concretio Silicea was referred to by several different names,such as"Zhu Huang""Tian Zhu Huang""Zhu Gao""Zhu Tang",and"Zhu Huang".The earliest known formal usage of the name"Tian Zhu Huang"was found in the book Ri Hua-zi's Materia Medica(Ri Hua Zi Ben Cao).Throughout various ancient texts,the earliest recorded information about Bambusae Concretio Silicea also appeared in Ri Hua-zi's Materia Medica,not in Materia Medica of Sichuan(Shu Ben Cao)or other ancient texts.Ri Hua-zi's Materia Medica provided relevant descriptions of its origin,medicinal properties,and indications,albeit with some errors due to limited knowledge.However,this has been a valuable starting point for future research on Bambusae Concretio Silicea and holds pioneering significance in forming a mature system.As the research delved deeper,the medicinal properties of Bambusae Concretio Silicea have been consistent since Ri Hua-zi's Materia Medica,and the understanding has gradually improved through years of clinical verification.During the investigation process,the authors found limited records on the quality evaluation of Bambusae Concretio Silicea in ancient texts.Although the information is scarce,it serves as a foundational basis for establishing corresponding quality grading standards for Bambusae Concretio Silicea in the future.

目的:建立并优化根癌农杆菌介导的甘草内生真菌粉红粘帚菌的遗传转化体系,优选粉红粘帚菌定殖甘草的条件,为生物菌肥的开发及优质甘草的育种奠定基础.方法:分别从乙酰丁香酮浓度、共培养时间和受体真菌分生孢子浓度3个方面对根癌农杆菌介导的遗传转化(ATMT)条件进行优化并优选转化子.采用正交试验,以共培养温度、共培养时间和孢子浓度为考察因素,定殖率为指标,优化粉红粘帚菌定殖甘草的条件.结果:粉红粘帚菌最适转化条件为共培养时间60 h、孢子浓度1×107cfu·mL-1、乙酰丁香酮浓度150μmol·L-1,此条件下的转化效率为每1×107个受体真菌孢子可获得135个转化子.通过标记基因绿色荧光蛋白(GFP)的克隆和p-葡萄糖苷酸酶(GUS)染色结果检测转化的准确性与稳定性.粉红粘帚菌通过浸种法定殖甘草的最佳条件为共培养温度25℃、共培养时间36 h、孢子浓度1×106cfu·mL-1,此条件下定殖率71.11％.结论:本研究成功建立了稳定高效的粉红粘帚菌ATMT体系及其定殖甘草的技术体系,可为甘草生物菌肥的开发奠定基础.

目的 研究金银花的表面微生物的组成和抑菌功能,一方面为金银花药材表面可能存在的污染菌提供参考,另一方面对其功能菌进行深入挖掘.方法 以河南封丘、山东平邑、河北巨鹿三个地区的金银花药材为研究对象,利用微生物培养技术,分离并鉴定其种属,并在此基础上检测其抑菌活性.结果 金银花表面菌群丰富,分离得到表面微生物53株,分布于5门7纲8目12科15属;群落组成显示,存在镰刀菌属、链格孢属、裂褶菌属、青霉属等污染菌;在抑菌拮抗实验中得到一株真菌,鉴定为非洲哈茨木霉,其能够抑制枯草芽孢杆菌.结论 金银花表面既存在污染菌,同时也存在功能菌,为防治真菌的污染和反向挖掘微生物的功能提供理论依据.

鸡血藤是传统的活血化瘀类中药,具有活血补血、调经止痛、舒筋活络的功效,临床应用广泛.约有26种药用植物曾作为鸡血藤在临床使用,其所含化学成分复杂多样,主要包括黄酮、蒽醌、木脂素、三萜及挥发油等,表现出促进造血功能、抗肿瘤、抗炎、抗氧化、保肝及抗人免疫缺陷病毒(HIV)等多种药理活性.根据本草著作、文献研究和现代应用情况,分别对来自豆科和木兰科的11种鸡血藤类药材的主要化学成分和药理活性进行综述,对比两者所含成分及药理活性的异同,为其进一步的开发与合理利用提供依据.

Heosemys grandis , a species of Asian water turtle, that has a wide range of applications in the food and pharmaceutical industries. Since some processed products cannot be identified exclusively by morphological and microscopic identification, a reliable and quick approach to guarantee authenticity is critical. Thus, we fostered an effective and stable molecular identification system to identify Heosemys grandis based on DNA mini-barcoding and species-specific primers PCR technology. A total of 48 turtle samples from 16 different species were collected. To distinguish Heosemys grandis from its counterfeits, DNA mini-barcoding and a pair of species-specific primers were designed and verified by PCR after analyzing the COI sequences of samples. The results showed that only Heosemys grandis samples could generate a single clear band following amplification using species-specific primers. Employing DNA mini-barcoding to amplify samples can verify authenticity by sequence alignment. These findings indicated that species-specific primers PCR technology combined with DNA mini-barcoding could accurately detect the authenticity of Heosemys grandis . This technology broadens the application of molecular biology techniques in the food and pharmaceutical industries. It provides a reliable and convenient method for identifying raw materials to standardize the market and protect customers’ rights and interests.

目的:研究不同产地柴胡气味差异及其气味形成的物质基础.方法:利用电子鼻和顶空气相色谱-质谱法(HS-GC-MS)检测来自山西、内蒙古及黑龙江3个地区18批柴胡样品的气味及具体挥发性成分,通过偏最小二乘法-判别分析(PLS-DA)分析不同产地柴胡样品-传感器响应值-挥发性成分之间的关系.结果:雷达图显示不同产地柴胡响应值存在差异,电子鼻的检测结果经主成分分析(PCA)后进行线性判别分析(LDA)能较好区分来自山西、内蒙古和黑龙江的柴胡样品.采用HS-GC-MS共检测出76个挥发性成分,主要包括醛类、芳香类、酮类、醇类等物质.通过SIMCA 14.0筛选变量投影重要性分析值(VIP)>1的挥发性成分,得出11种主要差异性物质,包括2-正戊基呋喃、α-蒎烯、β-蒎烯、庚醛、三乙胺、十一烷、对异丙基甲苯、2-正丁基呋喃、1-戊醇、己醛、2-辛酮.结论:利用挥发性物质的相对质量分数和电子鼻感应值做PLS分析,发现造成山西省与其他地区柴胡电子鼻检测气味差异的物质基础主要是己醛、1-戊醇、2-辛酮、2-正丁基呋喃.

The content of total flavonol glycosides in Ginkgo Folium in the planting bases was determined by high performance liquid chromatography(HPLC).The samples were extracted by reflux with methanol-25% hydrochloric acid.The HPLC conditions were as follows: Agilent ZORBAX SB-C_(18) column(4.6 mm×250 mm, 5 μm), isocratic elution with mobile phase of 0.4% phosphoric acid solution-methanol(45∶55), flow rate of 1 mL·min~(-1), column temperature of 30 ℃, detection wavelength of 360 nm, and injection vo-lume of 10 μL.A method for the determination of terpene lactones in Ginkgo Folium was established based on ultra-high performance liquid chromatograph-triple-quadrupole/linear ion-trap tandem mass spectrometry(UPLC-QTRAP-MS/MS).The UPLC conditions were as below: gradient elution with acetonitrile-0.1% formic acid, flow rate of 0.2 mL·min~(-1), column temperature of 30 ℃, sample chamber temperature of 10 ℃, and injection volume of 10 μL.The ESI~+and multiple reaction monitoring(MRM) were adopted for the MS.The above methods were used to determine the content of total flavonol glycosides and terpene lactones in 99 batches of Ginkgo Folium from 6 planting bases, and the results were statistically analyzed.The content of flavonoids and terpene lactones in Ginkgo Folium from different origins, from trees of different ages, harvested at different time, from trees of different genders, and processed with different methods was compared.The results showed that the content of total flavonol glucosides in 99 Ginkgo Folium samples ranged from 0.38% to 2.08%, and the total content of the four terpene lactones was in the range of 0.03%-0.87%.The method established in this study is simple and reliable, which can be used for the quantitative analysis of Ginkgo Folium.The research results lay a basis for the quality control of Ginkgo Folium.

针对中医药院校部分毕业生"知药名,不识药"的不良状况,北京中医药大学中药学院近年来实施中药专业学生中药饮片等级考试前置毕业措施.从考试对象、考试内容、考核方式及考试成绩等方面对该自主学习为导向的等级考试模式进行探讨,以期为优化饮片等级考试模式、培养高素质中药学人才提供参考.

BackgroundThe glycyrrhizic acid biosynthesis pathway does not exist in isolation, but is connected with the biosynthesis pathways of other secondary metabolites in licorice and finally forms a network. Our previous study found that exogenous spraying of appropriate concentration of abscisic acid (ABA) could increase the content of glycyrrhizic acid (GA) in licorice. However, the mechanism of action remains unknown. We aim to understand the molecular mechanism of ABA promoting the synthesis of GA in licorice and find the molecular marker for the high content of GA germplasm material.MethodsWe analyzed the expression of the key gene of β-AS for GA synthesis after applying ABA, the key functional genes NCED1 , NCED3 and NCED4 in the process of ABA synthesis were overexpression, and analyzed the relationship between the SNP polymorphism of the NCED1 , NCED3 , NCED4 and the content of the GA and ABA in 13 different provenances of licorice with the grey correlation analysis. ResultsThe appropriate concentration of ABA treatment could increase the content of the GA through improving the expression of β-AS . There were significant differences in the content of ABA and GA among the 13 provenances, and the 3 members of the NCEDs family of different provenances had abundant SNP variation sites. Grey correlation and overexpression of NCEDs function both showed that the effect of promoting the synthesis of ABA and GA: NCED1 437 bp G type > NCED3 966 bp G type > NCED4 845 bp A type. All of the above indicated that NCEDs gene variation was the reason for the diversity of GA and ABA content. When selecting high GA germplasm, more priority should be given to NCED1 gene 437 bp G type, NCED3 gene 966 bp G type and NCED4 gene 845 bp A type. ConclusionThis study provides a basis for the selection of excellent GA content germplasm of licorice, and provide some reference for producing a high quality cultivated licorice.

The major objective of this study was to evaluate the inhibitory effect of essential oil (EO) from Zanthoxylum armatum DC. on Aspergillus flavus. The chemical composition of the EO was identified by gas chromatography–mass spectrometer. The minimum inhibitory concentration (MIC) of EO was investigated by liquid fermentation. The morphology, colony number, and aflatoxin content of A. flavus in platycladi semen were investigated by stereomicroscopy, scanning electron microscopy, plate counting, and high-performance liquid chromatography. The results indicated that the MIC of EO was 0.8 μL⋅mL–1, and the main components were β-phellandrene (7.53%), D-limonene (13.24%), linalool (41.73%), terpinen-4-ol (5.33%), and trans-nerolidol (6.30%). After the EO fumigated the platycladi semen, the growth of A. flavus slowed, and the mycelium shrank considerably. The number of colonies after EO treatment at room temperature and cold storage was significantly reduced, the inhibition effect was better under cold storage, and the aflatoxin B1 content did not exceed the standard within 100 days. Therefore, this study demonstrated the good potential of A. flavus growth inhibition during the storage of platycladi semen.

目的:研究不同产地远志中黄曲霉毒素B1、B2、G1、G2(AFB1、AFB2、AFG1、AFG2)的污染情况及贮藏过程中黄曲霉菌的变化.方法:收集12批不同产地的远志药材,采用性状、显微及DNA分子鉴定方法对其表面的黄曲霉菌进行鉴定;利用超高效液相色谱-三重四级杆串联质谱法(UPLC-MS/MS)测定其黄曲霉毒素含量;通过稀释平板法观察远志在北京贮藏6个月以来其表面黄曲霉菌的变化情况.结果:12批远志药材表面均分离鉴定出了黄曲霉菌,且全部检出黄曲霉毒素,其中33.3％的样品超过《中华人民共和国药典》2020年版中远志项下黄曲霉毒素B1、黄曲霉毒素总量的限定标准.结合北京的气候条件分析,在温度、湿度较高的夏季,远志表面的黄曲霉菌数量最多,在温、湿度较低的秋冬季节,远志表面的黄曲霉菌数量较低.结论:远志表面黄曲霉毒素污染问题十分严重,贮藏条件的温度、湿度变化对远志表面的黄曲霉菌数量有很大影响,可作为优化远志贮藏条件的参考依据.

目的:研究不同生境甘草内生真菌的种类和差异.方法:通过分离、鉴定2种不同生境的甘草内生真菌,系统分析和比较了两者内生真菌的种类和多样性.采用组织块分离法分离甘草内生真菌,结合形态学、显微学和分子生物学技术鉴定内生真菌;利用MEGA 7.0软件构建系统进化树,对菌株进行聚类分析;运用SPSS 22.0版软件进行内生真菌的多样性分析.结果:共分离得到365株内生真菌,归属于4纲5目6科9属30种,其中214株来自野生生境的甘草,151株来自栽培生境的甘草;镰刀属Fusarium(62.62％)和曲霉属Aspergillus(12.15％)为野生生境甘草的优势菌属;镰刀属(62.91％)和土赤壳属Ilyonectria(12.58％)为栽培生境甘草的优势菌属;分离出11株野生生境甘草特有的内生真菌,如尖孢曲霉Aspergillus aculeatinus、枝状枝孢菌Cladosporium cladosporioides、粉红粘帚菌Clonostachys rosea等;野生生境甘草内生真菌的总分离率和多样性指数(97.27％和2.07)均高于栽培甘草(68.64％和1.46).结论:明确了2种不同生境甘草内生真菌的种类和多样性差异,丰富了甘草内生真菌菌种库,为进一步研究药用植物内生真菌对药材质量的影响提供了理论依据.

分别对3个不同版本《分子生药学》教材的整体结构内容和章节进行分析,通过总结其各自的优势,提出要结合各个版本教材的亮点,进一步增强将来新版本《分子生药学》教材的完整性、创新性和趣味性,以期为《分子生药学》教材内容的体系建设与发展提供依据.

目的:对比分析天竺黄和人工天竺黄的微观结构及元素组成,为天竺黄和人工天竺黄的快速鉴别提供参考.方法:采用普通光学显微镜与偏光显微镜比较天竺黄和人工天竺黄的粉末显微特征与偏光性,联用扫描电镜-能谱仪(SEM-EDS)比较天竺黄和人工天竺黄的微观形貌与元素组成.结果:普通光学显微镜显示,天竺黄与人工天竺黄的差异在于有色团块和条状物;偏光显微镜显示,天竺黄与人工天竺黄均无偏光性;扫描电镜显示,天竺黄呈粒状集合体、粗糙,而人工天竺黄紧实、平滑;能谱仪检测结果表明,天竺黄和人工天竺黄的主要元素含量顺序及其他元素种类有一定差异.结论:天竺黄和人工天竺黄的微观结构及元素组成存在差异性,综合利用普通光学显微镜、扫描电镜和能谱仪可快速鉴别天竺黄和人工天竺黄.