Tang Huan, Xiang Li, Li Xi-Wen, Sun Wei, Wang Mei-Na, Huang Yun-Feng, Ye Meng
College of Forestry, Sichuan Agricultural University, Chengdu 611130, China.
Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine, Institute of Chinese Materia Medica，China Academy of Chinese Medical Sciences, Beijing 100700, China.
Zhongguo Zhong Yao Za Zhi. 2017 Jun;42(11):2058-2067. doi: 10.19540/j.cnki.cjcmm.2017.0090.
In this study, DNA barcoding was used to validate the traditional morphological classification of medicinal plants of Orchidaceae. The 163 samples of 135 species belong to 49 genera which have been confirmed by morphological identification were collected. Candidate sequences, including matK, psbA-trnH and ITS2 sequences, were amplified, bidirectionally sequenced, and assembled. All the sequences were blasted to GenBank database at NCBI, then analyzed using Neighbor-joining tree method by MEGA 7.0. The results showed that the DNAs of 163 samples were successfully extracted. The amplification efficiency of matK, psbA-trnH and ITS2 sequences were 100%, 100% and 98.77%, respectively. The 487 sequences were obtained, 345 sequences of which have matched corresponding sequences in the GenBank database and 142 sequences were new sequences. The topology of NJ tree which were constructed with the matK sequences was better than the trees of psbA-trnH and ITS2 sequences. In conclusion, the matK, psbA-trnH and ITS2 sequences were complementary and suitable for identification of medicinal plants of Orchidaceae. DNA barcoding can be used as an auxiliary means for identification of medicinal plants of Orchidaceae.
在本研究中，采用DNA条形码技术对兰科药用植物的传统形态分类进行验证。收集了经形态学鉴定确认的49个属135种的163个样本。对包括matK、psbA-trnH和ITS2序列在内的候选序列进行扩增、双向测序和组装。将所有序列在NCBI的GenBank数据库中进行比对，然后用MEGA 7.0软件采用邻接法进行分析。结果表明，成功提取了163个样本的DNA。matK、psbA-trnH和ITS2序列的扩增效率分别为100%、100%和98.77%。共获得487条序列，其中345条序列在GenBank数据库中匹配到相应序列，142条为新序列。用matK序列构建的NJ树拓扑结构优于用psbA-trnH和ITS2序列构建的树。综上所述，matK、psbA-trnH和ITS2序列具有互补性，适用于兰科药用植物的鉴定。DNA条形码技术可作为兰科药用植物鉴定的辅助手段。
