藥材真偽之分子標誌及檢驗方法建立-蒲公英

袁秋英
行政院農業委員會 農業藥物毒物試驗所
中草藥於中國已流傳千年，超過一萬種植物具有療效，其中約10%為經常性使用的藥材。近年來中草藥藥材出現代用品及偽品，使用不當容易影響療效也可能發生中毒現象，這亦是中草藥品質管控不嚴謹之故。由於分生及PCR技術的研發，使得分子標誌普遍應用於植物物種之鑑定。DNA分子標誌法具有專一性、穩定性、微量、準確、易操作及快速等優點，其中ISSR方法為較新的技術，可用於鑑定族群的親緣關係及分辨品系間微小之差異。針對18S-26S rDNA序列之比對，亦為鑑定中草藥物種的有利證據，常被應用於被子植物之親源關係之研究。本研究利用ISSR技術及ITS-5.8S rRNA-ITS序列之差異，進而設計為PCR-RFLP及multiplex PCR檢測方法， 建立蒲公英及其偽品、替代品之檢測系統，提供為中草藥基原鑑定及品質控管之依據。
關鍵字：中草藥；分子標誌；基原鑑定

Molecular markers used in detection of adulteration species of Taraxacum mongolicum in Taiwan

Chiou-Ing Yuan
Taiwan Agricultural Chemicals and Toxic Substances Research Institute
Traditional China medicine (TCM) has been used for millennium in China. At present, over 10,000 species of plants have been found to have medicinal value and about 10% are frequently used in TCM. Substitutes and adulterants of TCM materials are often introduced intentionally or accidentally, thus serious interfering with their therapeutic effects, even leading to life- threatening poisoning. The absence of an objective and accurate inspection system is frequently cited as one of the major reasons for herbal poisoning. Benefiting from molecular cloning and PCR techniques, DNA markers have now become a popular means for identification and authentication of plant species. Inter simple sequence repeats (ISSR) are a new kind of molecular marker, can be used to identify different strains and races of organisms and to assess genetic relatedness in populations. Sequencing is a definitive means for identifying TCM. A representative sample is the internal transcribed spaces (ITS) from ribosomal DNA. The ITS region of 18S-26S rDNA has proves to be a useful sequence for phylogenetic studies in many angiosperm families. Novel methods for molecular authentication of Taraxacum mongolicum we’re established in this study, based on PCR-restriction fragment length polymorphism (RFLP) and direct sequencing of the ITS region of 18S-26S rDNA.
關鍵字：herbal medicine；molecular marker；authentication of medicinal plant