藏藥臭蒿及其近緣種藥材的ITS2,DNA條形碼鑒別

　　[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(tǒng)鑒定方法難以準(zhǔn)確快速鑒定。文章以ITS2序列為DNA條形碼，建立了一種快速準(zhǔn)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(gè)樣品，分別提取樣品DNA，對(duì)ITS2目標(biāo)序列進(jìn)行PCR擴(kuò)增、測(cè)序，獲得其ITS2序列信息；并從GenBank上下載同屬序列11條；運(yùn)用MEGA 6.0對(duì)所有32條ITS2序列進(jìn)行對(duì)比，計(jì)算種間種內(nèi)遺傳距離，構(gòu)建Neighbor Joining（NJ）系統(tǒng)進(jìn)化樹，并比較樣本ITS2序列間的二級(jí)結(jié)構(gòu)差異。結(jié)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(yuǎn)大于種內(nèi)最大距離，NJ樹顯示4種蒿屬植物種內(nèi)之間各單獨(dú)形成一個(gè)分支，ITS2序列二級(jí)結(jié)構(gòu)也顯示出差異。以ITS2序列作為DNA條形碼，可達(dá)到對(duì)臭蒿及其近緣蒿屬藏藥植物進(jìn)行準(zhǔn)確、快速的識(shí)別和鑒定的目的，準(zhǔn)確地弄清物種之間的系統(tǒng)進(jìn)化關(guān)系的目的，為該屬藥用植物的質(zhì)量控制、安全用藥及合理開發(fā)利用提供理論依據(jù)。
　　[關(guān)鍵詞] 臭蒿； 藏藥； DNA條形碼； ITS2； 二級(jí)結(jié)構(gòu)； 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(guān)熱詞搜索：條形碼 鑒別 藥材 藏藥 ITS2