DNA条形码等分子鉴定技术与动植物类中药材的鉴定

中药材鉴定是控制中药质量、确保用药安全与效果的首要环节。DNA分子鉴定是从基因层面上进行中药材鉴别的手段,准确率高。DNA分子鉴定包括电泳技术、免疫技术、随机扩增多态性DNA(RAPD)、限制性内切酶片段长度多态性(RFLP)、扩增片段长度多态性(AFLP)、简单重复序列(ISSR)及DNA条形码等,以DNA条形码应用最为广泛。DNA条形码是基因组中相对较短的、可用于物种鉴定的特异性基因片段。植物类中药材的条形码多选用核基因和叶绿体基因DNA片段,如叶绿体psbA-trnH基因、内转录间隔区(ITS)、叶绿体核酮糖-1,5-二磷酸羧化酶大亚基(rbcL)和RNA转录体Ⅱ型内含子剪切酶基因(matK)等;动物类中药材的条形码多来自核基因和线粒体基因DNA,主要有线粒体细胞色素C氧化酶亚基1(COⅠ)、核糖体RNA(rRNA)和线粒体细胞色素b基因(CytB)等。综述动植物类中药材DNA分子标记与鉴定技术应用进展,并探讨DNA条形码在药用动植物类中药材鉴定中存在的局限性及发展前景,为中药质量控制提供参考。

Application of DNA Barcoding and Other Molecular Techniques in Identification of Animal and Plant Traditional Chinese Medicine

The identification of traditional Chinese medicine(TCM)is the first step for quality control of traditional Chinese medicine.DNA molecular identification,is a method to identify TCM from the gene level by directly analyzing the genetic material polymorphism.It has high accuracy and is not easy affected by the development stage,tissue position,sample shape and external environmental factors.DNA molecular identification includes electrophoresis,immunoassay,random amplified polymorphic DNA(RAPD),restriction endonuclease fragment length polymorphism(RFLP),amplified fragment length polymorphism(AFLP),simple repeat sequence(ISSR)and DNA barcode,among which DNA barcode is most widely adopted.DNA barcoding is a relatively short specific gene segment in genome,which can be used for species identification.In the barcode of plant traditional Chinese medicine,nuclear gene and chloroplast gene DNA fragments are mostly selected,such as chloroplast psbA-trnH gene,internal transcribing spacer(ITS),chloroplast riboketose-1,5-diphosphate carboxylase large subunit(rbcL)and RNA transcriber typeⅡintron shear enzyme gene(matK).In animal traditional Chinese medicine,the bar code mostly comes from nuclear gene and mitochondrial gene DNA,mainly including mitochondrial DNA Cytochrome C oxidase subunit 1(COⅠ),ribosomal RNA(rRNA)and mitochondrial cytochrome b gene(CytB).DNA barcoding identification technology is one of the fastest developing methods in DNA molecular identification technology,which plays an increasingly important role in the identification of animal and plant TCM.In this paper,the application of DNA molecular marker and identification technology in animal and plant traditional Chinese medicine is reviewed.