基于中药材DNA条形码系统的泽泻种子鉴别研究  

Identification of Alisma plantago-aquatica Seeds Based on Traditional Chinese Medicine DNA Barcoding System

作　　者：张娜娜[1] 辛天怡[1] 金钺[1] 谷巍[2] 宋经元[1] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]南京中医药大学药学院,南京210046

出　　处：《世界科学技术-中医药现代化》2016年第1期18-23,共6页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委面上项目(81373922):基于ITS2互补碱基变化分析的中药基原物种鉴定研究;负责人:宋经元

摘　　要：目的:以泽泻种子为研究对象,利用中药材DNA条形码鉴定系统区分不同产地泽泻的基原物种。方法:采用植物基因组DNA提取试剂盒提取总DNA,扩增核内部转录间隔区ITS2序列并进行测序。利用软件CodonCode Aligner 6.0.2和MEGA 5.1对序列进行拼接,并用体式显微镜观察记录种子外观形态特征。结果:泽泻Alisma plantago-aquatica和东方泽泻Alisma orientale种子外观形态相似;两个物种的ITS2序列长度均为311 bp,GC含量为56.9%,ITS2序列在165 bp处存在T-A变异。该鉴定结果显示,20份四川产泽泻种子样本序列为泽泻A.plantago-aquatica,其他产区36份样本均为东方泽泻A.orientale。结论:中药材DNA条形码鉴定系统可为选择正确基原的泽泻种质提供指导,该系统在泽泻人工栽培生产中具有重要应用价值,可为其他中药材种子鉴定提供示范。This study was aimed to authenticate the Alisma plantago-aquatica seeds from different producing areas with the Traditional Chinese Medicine（TCM） DNA barcoding system.The total genomic DNA was extracted using plant DNA extraction kit.The internal transcribed spacer 2（ITS2） regions were amplified and sequenced.The sequences were assembled by the CodonCode Aligner 6.0.2 and analyzed by MEGA5.1.The morphological characteristics of seeds were observed using microscope.The results showed that the morphological characteristics of Alisma plantago-aquatica seeds and Alisma orientale seeds were similar.The ITS2 sequence lengths of both species were 311 bp.The GC content was 56.9%.One stable SNP（T-A） was detected at 165 bp in the ITS2 sequence.The identification results showed that 20 samples collected in Sichuan province were identified as A.plantago-aquatica,while 36 samples from other producing regions were A.orientale.It was concluded that the TCM DNA barcoding system can provide guidance for the accurate selection of Alisma plantago-aquatica seeds.This system had important application value in the cultivation of Alisma plantago-aquatica.It also provided demonstration for the identification of other TCM seeds.

关 键 词：DNA条形码 泽泻 东方泽泻 种子鉴定 ITS2 

分 类 号：R931.5[医药卫生—生药学]