[摘要] 臭蒿Artemisia hedinii有清熱涼血，退黃，消炎的功能，在藏藥中占有重要地位。蒿屬植物種類繁多，在青藏高原分布廣泛，且其外形較為相似，傳統鑒定方法難以準確快速鑒定。文章以ITS2序列為DNA條形碼，建立了一種快速準確鑒定臭蒿及其易混蒿屬藏藥植物的方法。從青藏高原采集蒿屬藏藥21個樣品，分別提取樣品DNA，對ITS2目標序列進行PCR擴增、測序，獲得其ITS2序列信息；并從GenBank上下載同屬序列11條；運用MEGA 6.0對所有32條ITS2序列進行對比，計算種間種內遺傳距離，構建Neighbor Joining（NJ）系統進化樹，并比較樣本ITS2序列間的二級結構差異。結果顯示：臭蒿、牛尾蒿、黃花蒿、艾蒿的遺傳距離較近，但四者種間最小距離均遠大于種內最大距離，NJ樹顯示4種蒿屬植物種內之間各單獨形成一個分支，ITS2序列二級結構也顯示出差異。以ITS2序列作為DNA條形碼，可達到對臭蒿及其近緣蒿屬藏藥植物進行準確、快速的識別和鑒定的目的，準確地弄清物種之間的系統進化關系的目的，為該屬藥用植物的質量控制、安全用藥及合理開發(fā)利用提供理論依據。
　　[關鍵詞] 臭蒿； 藏藥； DNA條形碼； ITS2； 二級結構； 鑒定； 資源利用
　　ITS2 sequence analysis and identification of medicinal Artemisia plants
　　REN Yaoyao1， JIANG Nanping1， LIU Ruiying1， SONG Liangke1， TAN Rui1*， GU Jian2*
　�。�1. School of Life Science and Engineering， Southwest Jiaotong University， Chengdu 610031， China；
　　2. School of Pharmacy， Southwest University for Nationalities， Chengdu 610041， China）
　　[Abstract] Artemisia hedinii occupies an important position in the Tibetan medicine. Plants in Artemisia vary a lot and are widely distributed in the QinghaiTibet Plateau， many plants in Artemisia look similar， making traditional identification methods laborious. In this article， ITS2 sequences were used as DNA barcoding to identify four kinds of confusable Tibetan medicine plants in Artemisia， aiming to establish a rapid and accurate identification methods. Twentyone samples in Artemisia were collected from the QinghaiTibet Plateau， ITS2 sequence PCR amplification and sequencing were conducted after the extraction of DNA. Another 11 sequence downloaded from Genbank were added to the analysis. Genetic distance calculation and analysis， building Neighbor Joining （NJ） phylogenetic tree were conducted by MEGA 6.0， also comparison of secondary structures of ITS2 sequences among samples. A. hedinii， A. annua， A. dubia and A. argyi shared close genetic distance， but the maximum distance between the four species was much greater than the minimum distance within each species， NJ tree showed that the four species went to four separate branches， differences among secondary structures of ITS2 sequences also made it clear to identify these medical plants. It could be an accurate and rapid method for identification and recognition， as well as the evolutionary relationships between the species by using ITS2 sequence as DNA barcode for plants of Tibetan Artemisia. The study provides theoretical basis for quality control， medication safety and rational exploitation.

相關熱詞搜索：條形碼 鑒別 藥材 藏藥 ITS2