首页|基于mini-barcode和ITS2联用方法的批量茵陈鉴定研究

基于mini-barcode和ITS2联用方法的批量茵陈鉴定研究

扫码查看
茵陈来源于菊科植物滨蒿Artemisia scoparia或茵陈蒿A。capillaris。为了科学准确地鉴定批量茵陈样品的基原及混伪情况,该研究采用了一种基于多种分子标记联合应用的综合方法。研究基于GenBank上茵陈蒿、滨蒿及其他常见蒿属植物的ITS2序列,使用MEGA软件构建系统发育树,评估ITS2序列鉴定效果,并通过组装茵陈蒿的叶绿体基因组,与已发表的茵陈蒿和滨蒿的叶绿体基因组进行比对,开发DNA微条形码(mini-barcode)。将ITS2序列与所得mini-barcode联用进行Sanger测序鉴定单株茵陈样品,结合DNA宏条形码技术鉴定批量茵陈样品。研究结果表明,ITS2序列能够区分中药茵陈与其他易混伪的同属物种,但无法区分茵陈蒿与滨蒿。基于叶绿体基因组的比对结果,选择rpoA—rps11区域作为mini-barcode的开发区域,并设计了能够准确区分茵陈蒿与滨蒿的特异性引物。最终,不同产地的单株代表性茵陈样品及批量茵陈样品的鉴定结果均为茵陈蒿。通过该研究所建立的ITS2与mini-barcode联合鉴定方法,为不同产地批量茵陈的准确鉴定提供了科学依据,为稳定的药材供应提供了支持。
Mini-barcode combined with ITS2 for identification of bulk Artemisiae Scopariae Herba
Artemisiae Scoporiae Herba is derived from Artemisia scoparia or A.capillaris.The accurate identification of the herbs,particularly when dealing with bulk samples,is critical for ensuring the quality and efficacy of the medicinal product.This study aimed to establish a comprehensive molecular approach by combining multiple markers for the precise identification of Artemisiae Scoporiae Herba.The ITS2 from A.scoparia,A.capillaris,and other common Artemisia species were retrieved from GenBank.MEGA was used to build a phylogenetic tree with these sequences,and the effectiveness of ITS2 in species identification was assessed.The analysis revealed that while ITS2 could distinguish Artemisiae Scoporiae Herba from other closely related species of Artemisia,it was insufficient to differentiate between A.scoparia and A.capillaris.To address this limitation,the chloroplast genome of A.capillaris was assembled and compared with the published chloroplast genomes of A.scoparia and A.capillaris,on the basis of which a DNA mini-barcode was developed.The rpoA—rps11 region was selected as the target for the development of mini-barcode due to its potential for distinguishing between these two species.Specific primers were designed to differentiate A.scoparia from A.capillaris.The ITS2 sequences and the newly developed mini-barcode were used together for Sanger sequencing to identify individual samples of Artemisiae Scoporiae Herba,while DNA metabarcoding was employed for the identification of bulk samples.The identification results of representative individual samples and bulk samples from different regions consistently confirmed A.capillaris.This study established a method that combined ITS2 and mini-barcode to identify bulk samples of Artemisiae Scoporiae Herba from different regions.This approach overcomes the limitations of morphological and chemical methods,enhancing species identification accuracy and supporting a stable supply of medicinal materials.

Artemisiae Scopariae Herbaidentification of original plantchloroplast genomemini-barcodemetabarcoding

李昕怡、郭华、马明雪、徐流巍、黄宇华、张昀、杨翠萍、何峰、田晓轩

展开 >

天津中医药大学现代中药创制全国重点实验室,天津 301617

贵州瑞和制药有限公司,贵州黔南 551200

贵州医科大学药学院,贵州贵阳 550004

茵陈 基原鉴定 叶绿体基因组 微条形码 宏条形码

2024

中国中药杂志
中国药学会

中国中药杂志

CSTPCD北大核心
影响因子:1.718
ISSN:1001-5302
年,卷(期):2024.49(24)