茅苍术菌核病病原菌的分离与鉴定

为明确茅苍术菌核病种类和系统发育学特征,从茅苍术种植基地采集感病植株进行病原菌分离、纯化,按照柯赫氏法则进行致病性验证及再分离培养,采用形态学观察及ITS、G3PDH和HPS603个基因核酸序列分析对所采集病原菌进行鉴定。结果表明,根据形态学和多基因系统发育分析,经致病性测定及现代分子鉴定,可以确定引起重庆茅苍术菌核病的病原菌为核盘菌(Sclerotinia sclerotiorum),且茅苍术是核盘菌的新寄主。本研究结果可为茅苍术菌核病防治提供参考。

基于瓶内-瓶外结合的濒危药用植物茅苍术组培快繁研究

目的:优化茅苍术离体培养方法及条件,建立一种茅苍术高效组培快繁技术体系。方法:以茅苍术无菌苗为外植体,优化培养条件,筛选最佳增殖培养基,并采用组培瓶内-瓶外结合对茅苍术继代芽苗进行生根诱导,获得组培苗。结果:茅苍术胚轴切割以横切、仅保留下胚轴最佳,平均增殖系数为3.80;光照强度6000 lx更适于茅苍术不定芽诱导,平均增殖系数达4.36。茅苍术最适不定芽增殖培养基为MS+6-BA 1.0 mg/L+NAA 0.5 mg/L+蔗糖50 g/L,增殖系数为12.96±1.55,单芽最高增殖系数可达15,农艺性状整齐度最高。瓶外生根过程中,育苗周期较瓶内生根方式可缩短14 d左右,最适生根基质为草炭∶珍珠岩∶蛭石=3∶1∶1;瓶外生根平均成活率达95.00%,显著高于瓶内生根。结论:通过结合瓶内培养与瓶外生根技术,建立了一种创新性的茅苍术组培快繁技术体系,在缩短育苗周期的同时,提高茅苍术成活率,为濒危药用植物茅苍术工厂化育苗提供科学可行的技术支持。

Ca^(2+)对茅苍术丛生芽诱导及生理特征的影响

为提高茅苍术组培苗品质,以信阳野生茅苍术种子获取的初代无菌试管苗为试材,采用植物组织培养试验方法,研究不同浓度Ca^(2+)对茅苍术丛生芽诱导及生理特征的影响。结果表明:Ca^(2+)浓度为1.50 mmol/L时茅苍术丛生芽增殖数最高,为16.36个;Ca^(2+)浓度为3.00 mmol/L时丛生芽总干重最高,为0.37 g;Ca^(2+)浓度为6.00 mmol/L时丛生芽总鲜重、单芽平均叶片数最高,分别为3.19 g、5.44个;Ca^(2+)浓度为12.00 mmol/L时茅苍术丛生芽的株高最高,为493.95 mm;Ca^(2+)浓度为24.00 mmol/L时叶绿素a含量、叶绿素b含量、类胡萝卜素含量、叶绿素总量和丛生芽SOD总活性最高,分别为1.54 mg/g、0.40 mg/g、0.36 mg/g、1.94 mg/g和329.36 U/g。在茅苍术组织培养过程中可根据不同需求选择相应的Ca^(2+)浓度。

三种苗前除草剂对茅苍术杂草防治效果及安全性评价

为了筛选出防效好、残留少且对环境影响较小的茅苍术苗前除草剂,选取常用的3种苗前除草剂乙草胺、乙氧氟草醚和二甲戊灵,研究其除草效果,对叶绿素荧光参数的影响,农药残留情况及其对土壤微生物真菌和细菌群落的影响。结果表明,乙草胺的除草效果较好,株防效和鲜重防效都达到了80%以上,对茅苍术叶绿素荧光参数影响较小,农残较少,对土壤细菌和真菌群落无显著性影响;乙氧氟草醚和二甲戊灵对叶绿素荧光参数有影响,短时间内有一定的土壤农残。综合防效和安全性,优选乙草胺作为茅苍术的苗前除草剂。

Optimization of Extraction Process of Fagopyri Dibotryis Rhizoma by Orthogonal Experiment

[Objectives]To optimize the water extraction process of Fagopyri Dibotryis Rhizoma.[Methods]The entropy weight method was used to determine the weight of epicatechin extraction rate and dry extract rate and calculate the comprehensive score.The water extraction process of Fagopyri Dibotryis Rhizoma was optimized by orthogonal design with the comprehensive score as the indicator and the amount of water,extraction time and extraction times as the factors.[Results]The optimum extraction process of Fagopyri Dibotryis Rhizoma was as follows:adding 10 times of water,extracting 3 times,and extracting for 60 min each time.[Conclusions]The optimized extraction process is stable and feasible,and can be used for the extraction of Fagopyri Dibotryis Rhizoma.

Fingerprint Study of Polygonati Rhizoma with Steaming and Exposing to the Sun Alternatively for Different Times

[Objectives]To explore the influence of different times of steaming and exposing to the sun on the fingerprint of Polygonati Rhizoma by studying the HPLC fingerprint of Polygonati Rhizoma processed products with different times of steaming and exposing to the sun,and to provide a basis for the determination of the best processing technology of Polygonati Rhizoma.[Methods]SETSAIL II AQ-C 18(5μm×250 mm×4.6 mm)was used as the column,the column temperature was 30℃,pure water(A)and acetonitrile(B)were eluted gradually,0-10 min,B(5%-10%),10-30 min,B(10%-35%),30-40 min,B(35%-60%),40-45 min,B(60%-100%),flow rate 1 mL/min,absorption wavelength 200 nm.[Results]The relative retained peak area RSDs of the common peaks in the precision,reproducibility and stability tests were all less than 5%.There were 17 common peaks in the fingerprint of nine batches of samples,and the retention time of Peak 2 was basically the same as that of the reference peak of 5-HMF.Peak 4 mainly existed in the chromatogram of Sample 3 to Sample 5,peaks 5 and 11 mainly existed after Sample 3,peaks 9,14 and 16 mainly existed after Sample 6,and peaks 12 and 17 mainly existed after Sample 4.[Conclusions]A total of 17 common peaks were obtained,and the Peak 2 was the designated peak,and the chemical components of each processed product were different.

The impact of^(60)Co-γirradiation on the chemical constituents of Chuanxiong Rhizoma

Background:In order to clarify the inmpat ofγirradiation on the chemical composition of traditional Chinese medicine,this paper carefully choosed Chuanxiong Rhizoma to carry on a demonstration study.Methods:Through a meticulous assessment,a comprehensive comparison was made between the irradiated and unirradiated Chuanxiong Rhizoma samples.The property characteristics were investigated by colorimeter and electronic nose.The changes in chemical structures and contents was analyzed by fourier infrared spectroscopy,high performance liquid chromatography and fingerprinting.In a quest to uncover the presence of any new radiolysis products,cutting-edge techniques like ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry and gas chromatography-mass spectrometry were employed.Moreover,the difference of antioxidant activity were investigated.Results:The irradiation doses within 12 kGy had no significant effects on the content of the main chemical components,characteristics and in vitro antioxidant activity of Chuanxiong Rhizoma,while changes in some functional groups and degradation of some volatile oil components containing olefins need further study.Conclusion:This study indicates that^(60)Co-γirradiation is a stable method for sterilization of Chuanxiong Rhizoma.It’s also provide a reference for the establishment of irradiation standards for Chuanxiong Rhizoma and other aromatic medicinal plants.

Exploring the mechanism of active ingredients of Smilacis Glabrae Rhizoma in treating migraine headache through network pharmacology and animal experiments

Background:To explore the potential mechanism of action of the active ingredients of Smilacis Glabrae Rhizoma(SGR)in the treatment of migraine using network pharmacology and in vivo experiments.Methods:Through the search of Traditional Chinese Medicines Systems Pharmacology Database and Analysis Platform,Genecards,Drugbank and other databases,we obtained active ingredients,targets of SGR and related disease targets of migraine,and took the intersection for protein-protein interactions analysis.After constructing the network diagram,network topology analysis was performed to derive the core targets and key active ingredients,and Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed.Finally,molecular docking was performed and validated by in vivo experiments.In vivo experiments,18 male BALB/c mice were selected,and the SGR group was fed with SGR drinking tablet concentrate,and nitroglycerin injection was used to construct a mouse model of migraine.Enzyme-linked immunosorbent assay test was used to detect the levels of TNF-α,IL-1β,IL-6,and AKT1 in plasma.Results:The results showed that the core targets of SGR for the treatment of migraine were TNF-α,IL-1β,IL-6,and AKT1.These core targets and key active ingredients had better binding ability.Compared with the blank group,the number of head scratching in the model group increased.Compared with the model group,there was a significant reduction of the number of head scratching in the SGR group.In comparison with the blank group,the protein level in the plasma in the model group was markedly higher.Compared with the model group,the protein level in the SGR group was significantly lower.Conclusion:SGR has the characteristics of improving migraine based on multi-targets,multi-components and multi-pathways,and the mechanism of action may be related to the inhibition of the release of inflammatory factors,neuron protection,and interference with apoptosis and other processes.

麸炒茅苍术水提物对LPS诱导人结肠上皮细胞炎症损伤的保护作用

目的考察麸炒茅苍术水提物对脂多糖(LPS)诱导正常人结肠上皮细胞(HCoEpiC)炎症损伤的保护作用。方法以脂多糖复制人结肠上皮细胞炎症模型,MTT法考察HCoEpiC细胞存活率,ELISA法检测细胞上清液IL-1β、IL-8、IL-12、IL-4、IL-6、TNF-α水平,RT-qPCR法检测细胞IKKα、NF-κB、TNF-α、IL-4、IL-8、IL-6 mRNA表达,Western blot法检测IL-8、IL-6、NF-κB、IKKα、p-IKKα、p-IKKβ蛋白表达。结果茅苍术生品和麸炒品水提物均能提高HCoEpiC细胞存活率(P<0.05),升高IL-4水平和mRNA表达(P<0.05),降低IL-6、IL-8、IL-1β、IL-12、TNF-α水平,下调IL-6、IL-8、TNF-α、NF-κB mRNA表达及IL-6、IL-8、p-IKKα、p-IKKβ、NF-κB蛋白表达(P<0.05),并且麸炒品作用优于生品。结论茅苍术水提物具有抗脂多糖诱导人结肠上皮细胞炎症损伤的作用,其机制可能与调控IKK/NF-κB信号通路有关,麸炒后该作用有增强的趋势。

幼龄核桃林下套种茅苍术高效种植技术初探

针对十堰地区核桃种植面积大、投入周期长及单纯种植核桃前几年经济效益低的问题,根据核桃和茅苍术的生物学特性,通过长期实践,总结出一种幼龄核桃林下套种茅苍术的高效栽培模式,不仅提高了土地利用率,而且经济效益较高,以期能为种植户提供参考。

茅苍术AlPMK1、AlPMK2基因的克隆和表达分析

目的从茅苍术中克隆萜类化合物生物合成途径中的甲羟戊酸-5-磷酸激酶(PMK)基因,并对其进行序列分析、原核表达及基因表达分析。方法基于茅苍术转录组数据,采用RT-qPCR法克隆获得茅苍术的2条甲羟戊酸-5-磷酸激酶基因的全长序列,命名为AlPMK1、AlPMK2,使用生物信息学软件预测AlPMK1,AlPMK2编码蛋白的特征;利用MEGA 6.06软件构建系统进化树;构建原核表达载体,在大肠杆菌中诱导表达;运用RT-qPCR检测该基因在不同产地茅苍术中的相对表达及经不同时间茉莉酸甲酯诱导后的表达情况。结果AlPMK1、AlPMK2基因的ORF长度分别为1512、1590 bp,分别编码503 aa、529 aa。2个基因的二级结构中α螺旋和无规则卷曲占比最高,均属不稳定的酸性蛋白;结构功能域分析证明AlPMK1、AlPMK2分别在4~490 aa,40~527 aa处包含甲羟戊酸激酶的保守结构域;跨膜结构域分析预测,两者均为膜外蛋白;SDS-PAGE结果显示,分别在55、58 kDa处有清晰的目的蛋白表达。基因表达结果显示,在不同时间的茉莉酸甲酯诱导下AlPMK1、AlPMK2的基因表达分别在48、24 h达到最高值,且均在3 h表达最低。不同产地的茅苍术AlPMK1、AlPMK2基因具有组织特异性,两者均在岳西茅苍术的叶中表达量最高。结论本研究首次克隆2条茅苍术AlPMK1、AlPMK2基因,并对其进行表达分析,为进一步探究茅苍术萜类生物合成途径奠定基础。

基于UPLC-MS/MS的茅苍术地上部分黄酮类成分分析研究

试验采用超高效液相色谱仪-轨道阱高分辨率质谱仪(UPLC-MS/MS)对茅苍术地上部分中黄酮类成分进行定性、定量分析。结果显示,在正离子模式下共检测出19个黄酮类物质(10个黄酮、5个异黄酮、2个花色素、1个黄烷醇和1个二氢黄酮),不同部位鉴定成分数量顺序为:苍术花(18个)>苍术茎(16个)>苍术叶(8个);在负离子模式下检测出23个黄酮类物质(13个黄酮、5个异黄酮、3个黄烷醇、1个花色素和1个二氢黄酮),不同部位鉴定成分数量无明显差别,分别为苍术花(12个)、苍术茎(10个)和苍术叶(15个);此外,黄烷醇类几乎只在苍术叶部位被检测到,且为苍术叶中主要黄酮类成分,(+)-儿茶素和表儿茶素没食子酸酯的占比高达16.68%、3.65%。研究表明,茅苍术地上部分黄酮类成分种类较多,含量较为丰富,有较大的开发潜力,可对目标成分进行提取和进一步的活性研究,以期将其开发为动物饲料添加剂,从而实现茅苍术地上部分资源的高效利用。

茅苍术的全长转录组分析

为探究茅苍术Atractylodes lancea(Thunb.)DC.中药用活性成分生物合成的遗传基础,对茅苍术进行三代全长转录组测序,通过生物学信息学分析,构建了茅苍术全长转录本数据库.共获得26238条平均长度为1360 bp的全长转录本,其中25413条(96.86%)在NR、KEGG、GO、KOG和Swiss-Prot 5个数据库中得到注释.根据NR比对结果,茅苍术与菊科菜蓟属刺苞菜蓟(Cynara cardunculus)最为相似;根据GO注释分析,8371条转录本注释到生物学过程、分子功能和细胞组成的三大类共48条GO条目;有15848条转录本注释到KOG数据库,参与基本功能预测序列数目最多,为2598条;在KEGG注释分析中,共有12137条转录本被注释到130条代谢通路;基因结构分析中,得到325条LncRNA、24723条CDS序列,检测到SSR位点8027个;根据转录因子分析,共预测到81个TF家族,包括1830个转录因子.该研究获得了茅苍术的全长转录组数据库,并为后续开展苍术的育种、活性成分的生物合成机制以及遗传多样性等分子研究提供宝贵的遗传资源.

不同氮素形态对茅苍术光合特性和光合氮素利用效率的影响

研究不同氮素形态对茅苍术光合特性和氮素吸收的影响,为其栽培氮肥高效利用提供理论依据。以一年生茅苍术为材料,研究不施氮肥、施用硝态氮、铵态氮和酰胺态氮对茅苍术叶片光响应曲线和光合氮素利用效率的影响。结果表明,施用氮肥可不同程度改善叶片光合特性,其中硝态氮处理的叶绿素含量、表观光量子效率、最大净光合速率,光饱和点、净光合速率、气孔导度、蒸腾速率均最高,但其光补偿点和胞间二氧化碳浓度显著低于铵态氮和酰胺态氮处理。同时,硝态氮处理能显著增大叶面积、降低比叶重,促进植株生长,使得其整株生物量比铵态氮和酰胺态氮处理提高6.89%和17.05%。此外,硝态氮处理还增加叶片氮素含量,提高光合氮素利用效率,分别比铵态氮和酰胺态氮处理提高2.43%和6.76%。可见,茅苍术光合特性对硝态氮更敏感,施用硝态氮肥能改善光能特性,促进氮素高效利用。

大别山区域茅苍术生境与仿野生栽培

结合调研情况阐述了大别山苍术生境要素,提出了苍术仿野生栽培的见解,倡导苍术仿野生栽培需把握产地近似、共生共长、生态环保、健身管理等原则与理念,总结了林下、梯带、林药、茶药、黍药、药药等多种较为成功的仿野生栽培模式。

不同覆盖物对大别山茅苍术出苗的影响

为了解大别山区域茅苍术[Atractylodes lancea(Thunb.)DC.]育苗过程中不同覆盖水平对茅苍术苗出苗与生长的影响,设置不同厚度地膜覆盖、松针覆盖、稻稃覆盖、杂灌叶覆盖与菌杂混土等处理。结果表明,不同覆盖物对茅苍术苗出苗数量、生长势、块茎鲜重、根长及健株率等性状的影响明显,其中地膜覆盖、菌杂混土处理的出苗质量显著优于无覆盖处理,稻稃覆盖、杂灌叶覆盖处理显著差于无覆盖处理,松针覆盖处理略优于对照,但未达显著水平。除健株率与成苗数量相关性未达到显著水平外,成苗数量、生长势、块茎鲜重、根长、健株率等性状两两之间呈显著正相关。不同覆盖物处理田块的杂草群相也有所不同,部分杂草如苍耳、问荆、蒿等在遮光覆盖条件下发生早而快。适合的土壤类型、良好的土壤墒情与微生态环境、适宜的温度与光照条件,均有利于促进茅苍术苗早生快发。根据茅苍术苗的成苗情况、生长势与整齐度基本能够判断幼苗的性状与全株生长状况。

茅苍术茎、叶、花中多类型资源性化学成分的分析与评价

目的:分析与评价茅苍术茎、叶、花中多类型资源性化学成分,为其资源化利用提供数据支撑。方法:采用紫外—可见分光光度法对茅苍术茎、叶、花中的总黄酮、总酚类、总蛋白质进行分析和评价。结果:总黄酮、总酚类在茅苍术地上部分不同部位中含量变化具有相似规律,即茅苍术花中含量>茅苍术叶中含量>茅苍术茎中含量。其中,茅苍术花中总黄酮和总酚类分别高达(24.01±0.16)mg/g和(0.71±0.02)mg/g。3个部位的总蛋白含量均较高,其中叶中含量最高,为(45.98±1.16)mg/g;其次花中含量为(43.94±0.98)mg/g;茎中含量最少,为(36.20±0.67)mg/g。从资源性成分总体分布情况来看,茅苍术茎中所含的总黄酮、总酚类和总蛋白量都最少。结论:茅苍术地上部分含有丰富的总黄酮、总酚和总蛋白等资源性化学成分,尤其是茅苍术花部位,不同部位之间各类型成分的含量差异较为明显。研究结果为茅苍术茎、叶和花部位的精细化利用与产业化开发提供科学依据。

特征图谱结合主成分分析研究生、麸茅苍术标准汤剂质量差异

目的:建立生、麸茅苍术标准汤剂特征图谱,并分析2种饮片标准汤剂的质量差异。方法:采用UPLC方法建立13批生、麸茅苍术标准汤剂特征图谱,结合主成分分析寻找区分2种标准汤剂质量差异的共有峰,统计分析共有峰之间比值,探讨2种标准汤剂的质量差异。结果:建立了生、麸茅苍术标准汤剂的特征图谱,共确定13个共有峰,指认了4个成分;通过对比特征图谱找出了差异峰。主成分分析将2种标准汤剂分别聚为Ⅰ类,2、10号色谱峰的峰面积比值可区分2种标准汤剂。结论:生、麸茅苍术标准汤剂质量存在差异,特征图谱与主成分分析能有效进行区分,差异峰峰面积比值可作为其质量专属性鉴别指标,为后期生、麸茅苍术配方颗粒的质量控制提供有效判别模式。

Experimental Study of Emilia Sonchifolia Combined with Coptidis Rhizoma for the Prevention and Treatment of Oral Ulcer

Objective: This paper aims to investigate the therapeutic effect of the combination of Emilia Sonchifolia and Coptidis Rhizoma on oral ulcer rats. Methods: 36 SD rats of half male and half female were kept for 7 days, and 6 rats among them were selected as normal group by random sampling method, and the rest rats were randomly divided into model group, positive control group, Emilia Sonchifolia group, Coptidis Rhizoma group and combined group after the establishment of oral ulcer model. The normal group and model group were given blank film, the positive control group was given Guilin Watermelon Frost, and the Emilia Sonchifolia group, Coptidis Rhizoma group and combined group were given the corresponding oral film, which was administered to the ulcer for 7 days continuously, 2 times per day. The healing of oral ulcer was assessed at the end of the last day of administration, and the ulcer area was calculated on the 1st, 3rd, 5th and 7th days after successful modeling, and the serum levels of IL-2 and TNF-α in rats were detected by Emilia Sonchifolia. Results: The grading of ulcer healing in the positive control group and the combined group was better than the other groups, and the difference was statistically significant when compared with the model group (p Coptidis Rhizoma group, Emilia Sonchifolia group, combined group and positive control group was smaller than that in the model group, and the difference was statistically significant (p Emilia Sonchifolia group, Coptidis Rhizoma group and combined group were all effective in reducing the area of oral ulcer, among which the combined group was more effective. The level of pro-inflammatory factor TNF-α was reduced and the level of anti-inflammatory factor IL-2 was increased in the Emilia Sonchifolia group, Coptidis Rhizoma group and combined group, and the difference was statistically significant (p Conclusion: The combination of Coptidis Rhizoma and Emilia Sonchifolia is effective in the treatment of oral ulcer in rats, and the effect of the combination is better than that of the drug alone.

Rhizoma paridis saponins protected against liver injury in diethylnitrosamine-induced mice

Background:Diethylnitrosamine,one of food additives,possessed a strong carcinogenic effect in human.Rhizoma paridis saponins,as the main active components of Paris polyphylla,have a good anti-cancer effect in our previous research.To verify their inhibitory effect on diethylnitrosamine-induced liver cancer,we carried out this study.Methods:We established diethylnitrosamine-induced mouse hepatocarcinoma models to evaluate antitumor of Rhizoma paridis saponins.Subsequently,gas chromatography-mass spectrometry was applied to analyze the metabolites in the urine and serum samples.Results:Rhizoma paridis saponins alleviated diethylnitrosamine-induced hepatocarcinogenesis.On the one hand,Rhizoma paridis saponins down-regulated the levels of liver function markers,such as alanine aminotransferase,aspartate transaminase and alpha fetoprotein.On the other hand,Rhizoma paridis saponins reduced metabolic disorders by increasing fructose and mannose metabolism,and decreasing pentose and glucuronate interconversion,inositol phosphate metabolism,and the process of saturated fatty acids transforming to unsaturated fatty acids,which based on the regulating mRNA expression of glucose transporter type 4,lactate dehydrogenase A,fatty acid synthetas,acetyl-CoA carboxylase and apolipoprotein A-I.Conclusion:Rhizoma paridis saponins has the potential application to inhibit chemical-induced hepatocarcinogenesis in the future.