世界科学技术-中医药现代化
世界科學技術-中醫藥現代化
세계과학기술-중의약현대화
WORLD SCIENCE AND TECHNOLOGY-MODERNIZATION OF TRADITIONAL CHINESE MEDICINE
2014年
8期
1730-1734
,共5页
宋明%张婉冰%张雅琴%孙伟%向丽%石晋丽%马孝熙%王俊%樊佳佳%张晓存%刘霞
宋明%張婉冰%張雅琴%孫偉%嚮麗%石晉麗%馬孝熙%王俊%樊佳佳%張曉存%劉霞
송명%장완빙%장아금%손위%향려%석진려%마효희%왕준%번가가%장효존%류하
金铁锁%ITS2%鉴定%混伪品
金鐵鎖%ITS2%鑒定%混偽品
금철쇄%ITS2%감정%혼위품
Psammosilene Radix%ITS2%identification%adulterants
目的：本研究利用ITS2序列对苗药金铁锁及其混伪品进行鉴定研究，从而确保药材质量，保障临床用药安全。方法：提取金铁锁药材及其混伪品基因组DNA，PCR扩增并双向测序获得ITS2序列。所有序列经过CodonCode Aligner V3.7.1拼接后，采用MEGA5.1软件进行序列比对，计算种内和种间Kimura 2-Parameter（K2P）遗传距离，并构建系统发育树。采用相似性搜索法、最近距离法、邻接（NJ）树法对ITS2序列鉴定能力进行评估。结果：金铁锁药材的ITS2序列长度为229 bp ，其ITS2序列种内最大K2P遗传距离小于与混伪品的最小种间K2P遗传距离；应用相似性搜索法表明ITS2序列能够准确鉴定金铁锁药材及其混伪品；基于ITS2序列构建NJ树，金铁锁及其混伪品均表现出良好单系性，均能明显区分。结论：ITS2序列能够稳定、准确地鉴定金铁锁药材及其混伪品，DNA条形码技术可为金铁锁药材及其混伪品的鉴定提供新的方法。
目的：本研究利用ITS2序列對苗藥金鐵鎖及其混偽品進行鑒定研究，從而確保藥材質量，保障臨床用藥安全。方法：提取金鐵鎖藥材及其混偽品基因組DNA，PCR擴增併雙嚮測序穫得ITS2序列。所有序列經過CodonCode Aligner V3.7.1拼接後，採用MEGA5.1軟件進行序列比對，計算種內和種間Kimura 2-Parameter（K2P）遺傳距離，併構建繫統髮育樹。採用相似性搜索法、最近距離法、鄰接（NJ）樹法對ITS2序列鑒定能力進行評估。結果：金鐵鎖藥材的ITS2序列長度為229 bp ，其ITS2序列種內最大K2P遺傳距離小于與混偽品的最小種間K2P遺傳距離；應用相似性搜索法錶明ITS2序列能夠準確鑒定金鐵鎖藥材及其混偽品；基于ITS2序列構建NJ樹，金鐵鎖及其混偽品均錶現齣良好單繫性，均能明顯區分。結論：ITS2序列能夠穩定、準確地鑒定金鐵鎖藥材及其混偽品，DNA條形碼技術可為金鐵鎖藥材及其混偽品的鑒定提供新的方法。
목적：본연구이용ITS2서렬대묘약금철쇄급기혼위품진행감정연구，종이학보약재질량，보장림상용약안전。방법：제취금철쇄약재급기혼위품기인조DNA，PCR확증병쌍향측서획득ITS2서렬。소유서렬경과CodonCode Aligner V3.7.1병접후，채용MEGA5.1연건진행서렬비대，계산충내화충간Kimura 2-Parameter（K2P）유전거리，병구건계통발육수。채용상사성수색법、최근거리법、린접（NJ）수법대ITS2서렬감정능력진행평고。결과：금철쇄약재적ITS2서렬장도위229 bp ，기ITS2서렬충내최대K2P유전거리소우여혼위품적최소충간K2P유전거리；응용상사성수색법표명ITS2서렬능구준학감정금철쇄약재급기혼위품；기우ITS2서렬구건NJ수，금철쇄급기혼위품균표현출량호단계성，균능명현구분。결론：ITS2서렬능구은정、준학지감정금철쇄약재급기혼위품，DNA조형마기술가위금철쇄약재급기혼위품적감정제공신적방법。
Psammosilene Radix is a famous miao national herb and it is rare and endangered nowadays. However, it is often confused with the root of Silene viscidula. In this study, the ITS2 regions was used to identify Psam-mosilene Radix and its adulterants. All DNA samples of Psammosilene Radix and its adulterants were extracted. The ITS2 regions were amplified and sequenced, and the final sequences were assembled using the CondonCode Aligner. The genetic distances were calculated by kimura 2-parameter (K2P) model and the Neighbor-Joining(NJ) phylogenetic trees were constructed using MEGA5.1.BLAST 1, the nearest distance and phylogenetic tree (NJ-tree)methods were used to assess the identification efficiency of the ITS2 region. Results indicated that the length of ITS2 sequences of Psammosilene Radix w 229 bp, the identification effficiency of ITS2 region using BLAST 1 was 100%;and the maximum intra-specific K2P distance were lower than the minimum inter-specific K2P distance. Additionally, the NJ tree based on ITS2 sequence indicated that Psammosilene Radix and its adul-terants could be distinguished clearly . In conclusion , the ITS2 region as DNA barcodes could identify Psammosi-lene Radix and its adulterants stably and accurately. Furthermore, the application of ITS2 barcode in the identifi-cation of TCM has a good prospective .
