云南丽江产粗茎秦艽溯源及道地药材初加工方法评价  

DNA barcoding of Gentiana crassicaulis in Lijiang and evaluation of root processing methods

作　　者：季文静 张玉萱[1] 赵志礼[1] 倪梁红[1] 李尉涛 竺琛欣 陈翔 杨少华[3] JI Wen-jing;ZHANG Yu-xuan;ZHAO Zhi-li;NI Liang-hong;LI Wei-tao;ZHU Chen-xin;CHEN Xiang;YANG Shao-hua(Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Kangqiao Chinese Herbal Pieces Co.,Ltd.,Shanghai 201315,China;Institute of Alpine Economic Plants,Yunnan Academy of Agricultural Sciences,Lijiang 674100,China)

机构地区：[1]上海中医药大学,上海201203 [2]上海康桥中药饮片有限公司,上海201315 [3]云南省农业科学院高山经济植物研究所,云南丽江674100

出　　处：《药学学报》2022年第2期507-513,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金面上项目(82073959,81173654);上海市卫生健康委员会中医药传承和科技创新项目(ZYCC2019012)。

摘　　要：种质来源、药材产地及其产出药材的加工炮制方法,是临床优质药材(饮片)生产的基本要素。秦艽为多来源品种,云南丽江为其基原之一——粗茎秦艽Gentiana crassicaulis道地药材产区。在药材传统栽培区确定试验样地,对所产药材进行加工炮制处理,即分别在产地、饮片厂进行平行初加工,分为切片或不切片(原个子)两大类,发汗处理或不经发汗处理,干燥方式为自然晾晒或加热烘干。分别测定其指标性成分,构建HPLC指纹图谱;同时,在课题组前期物种分类学鉴定及遗传背景分析基础上,筛选DNA片段,测定各加工炮制组药材样品以及丽江产野生植株的相关序列,以构建云南丽江道地产区药材的DNA条形码。结果显示:(1)试验样地样品及野生植株均为龙胆科龙胆属粗茎秦艽G.crassicaulis;(2)共得到270条trnC-GCA-petN、atpB-rbcL、psbN、ndhB-rps7及ycf1相关序列,丽江产栽培品分3个基因型,其中Ⅲ型与野生居群共享,由此构建云南丽江产粗茎秦艽(野生与栽培品)的DNA条形码;(3)各组马钱苷酸、龙胆苦苷的总含量均大于《中国药典》规定的2.5%;(4)HPLC指纹图谱中,确定9个共有峰,各组内相似度均大于0.999;(5)PCA得分图中,所有切片样品聚为一簇;而原个子样品分布较为分散。本工作可为生产临床质量稳定且优的秦艽药材(饮片)、多来源中药品种的科学评价及道地药材溯源及生产等提供科学资料。The key factors for producing the best quality Chinese herbal medicines are high-quality germplasm,suitable cultivation area and the proper processing methods for herbal raw materials.Gentiana crassicaulis in Gentiana(Sect.Cruciata),Gentianaceae is one of the original plants of the Chinese herb Qinjiao(Gentianae Macrophyllae Radix),and its type specimen was collected in Lijiang,Yunnan.There is a long planting history of the herb in this area.In this study a sampling plot was designated in these traditional planting areas.G.crassicaulis was planted and herbal raw materials were harvested from the plot.The raw materials were prepared locally and at a pharmaceutical factory in Shanghai using processing methods such as"sweating"or"no sweating","slicing"or"no slicing"(whole root),and"stoving"or"no stoving"(air drying).The quality of all processed samples was evaluated.In addition,molecular markers were determined for identifying cultivated and wild samples from Lijiang,Yunnan.The results are as follows:(1)Samples from the sampling plot and the field are taxonomically identified as Gentiana crassicaulis.(2)A total of 270 sequences of trnC-GCA-petN,atpB-rbcL,psbN,ndhB-rps7 and ycf1 were obtained,and three genotypes were determined from the cultivated samples;the type Ⅲ was shared by both cultivated and wild plants.Based on the molecular markers,a DNA barcoding method to identify cultivated and wild samples of G.crassicaulis from Lijiang,Yunnan was established.(3)Total content of loganic acid and gentiopicroside in all samples was ≥2.5%,and above the Chinese Pharmacopoeia(2020)limit.(4)In HPLC fingerprinting,9 common peaks were assigned and similarity between all samples was>0.999;and(5)In a PCA score plot all slice samples were clustered,while whole root samples were scattered.Therefore,our studies could provide basic data for optimizing the processing method,producing best quality Gentianae Macrophyllae Radix,and evaluating the quality of different ecotype varieties and the multiple origin of herbal medicines.

关 键 词：粗茎秦艽 道地药材 加工方法 DNA条形码 HPLC指纹谱 

分 类 号：R931[医药卫生—生药学]