中药女贞子的分子鉴定  

Species identification of Ligustrum lucidum

作　　者：王钰双 靳媛茜 刘康佳 郭畅 汪奕衡 徐超[3] 张志翔[1] 董文攀 WANG Yu-shuang;JIN Yuan-xi;LIU Kang-jia;GUO Chang;WANG Yi-heng;XU Chao;ZHANG Zhi-xiang;DONG Wen-pan(School of Ecology and Nature Conservation,Beijing Forestry University,Beijing 100083,China;State Key Laboratory Breeding Base of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;State Key Laboratory of Systematic and Evolutionary Botany,Institute of Botany,Chinese Academy of Sciences,Beijing 100093,China)

机构地区：[1]北京林业大学生态与自然保护学院,北京100083 [2]中国中医科学院中药资源中心道地药材国家重点实验室培育基地,北京100700 [3]中国科学院植物研究所系统与进化植物学国家重点实验室,北京100093

出　　处：《中国中药杂志》2023年第11期2940-2948,共9页China Journal of Chinese Materia Medica

基　　金：中国中医科学院科技创新工程项目(CI2021A03909);国家科技基础性工作专项(2021FY100200)。

摘　　要：女贞Ligustrum lucidum是木犀科Oleaceae女贞属Ligustrum多年生木本植物,其干燥果实女贞子是重要的中药。为了快速准确地进行女贞的分子鉴定,该研究评估了女贞属物种的3个专一DNA条形码片段(rbcL-accD、ycf1a、ycf1b)和4个通用DNA条形码(matK、rbcL、trnH-psbA、ITS2)的变异性和物种鉴定效率。结果显示,条形码matK、rbcL、trnH-psbA、ITS2、ycf1a的物种鉴定效率较低,rbcL-accD序列中有大量的插入和缺失序列,不适合作为专一条形码进行开发,ycf1b基因中的一段ycf1b-2条形码的PCR扩增和测序成功率高,且存在明显的DNA barcoding gap,是鉴定女贞的最优DNA条形码,可以实现女贞子的精准鉴定。此外,为进行DNA提取实验的优化,该文对女贞子果实的外果皮和中果皮、内果皮、种子3个部分进行了DNA的提取与检测结果分析,结果显示女贞子最适的DNA提取部位是种子,可以获得浓度高、质量优的DNA,满足物种鉴定的需求。该研究优化了女贞子DNA提取的实验技术,确定种子是最优的DNA提取部位;ycf1b-2为女贞子鉴定的专一条形码,实现了女贞子的快速精准鉴定,为市场监管奠定了基础。Ligustrum lucidum is a woody perennial plant of genus Ligustrum in family Oleaceae.Its dried fruit has high medicinal value.In this study,the authors evaluated the variability and species identification efficiency of three specific DAN barcodes(rbcLaccD,ycf1a,ycf1b)and four general DAN barcodes(matK,rbcL,trnH-psbA,ITS2)for a rapid and accurate molecular identification of Ligustrum species.The results revealed that matK,rbcL,trnH-psbA,ITS2 and ycf1a were inefficient for identifying the Ligustrum species,and a large number of insertions and deletions were observed in rbcL-accD sequence,which was thus unsuitable for development as specific barcode.The ycf1b-2 barcode had DNA barcoding gap and high success rate of PCR amplification and DNA sequencing,which was the most suitable DNA barcode for L.lucidum identification and achieved an accurate result.In addition,to optimize the DNA extraction experiment,the authors extracted and analyzed the DNA of the exocarp,mesocarp,endocarp and seed of L.lucidum fruit.It was found that seed was the most effective part for DNA extraction,where DNAs of high concentration and quality were obtained,meeting the needs of species identification.In this study,the experimental method for DNA extraction of L.lucidum was optimized,and the seed was determined as the optimal part for DNA extraction and ycf1b-2 was the specific DNA barcode for L.lucidum identification.This study laid a foundation for the market regulation of L.lucidum.

关 键 词：女贞子 DNA条形码 DNA提取 物种鉴定 

分 类 号：R282.5[医药卫生—中药学]