基于ITS条形码的云南丹参及其近缘种鉴定研究

Identification of Salvia yunnanensis and Relative Species Based on ITS Barcode

选取来自8个产地的材料,进行DNA提取、PCR扩增、测序。通过ITS条形码及其二级结构对云南丹参和正品药用丹参进行鉴定和聚类分析。采用DNAMAN 8.0进行碱基统计和序列比对;应用软件MEGA 6.0构建NJ系统进化树;并应用软件RNAstruct 6.0对各样品序列的ITS1和ITS2进行二级结构预测。结果表明:云南丹参和正品药用丹参ITS序列之间存在明显的差异,在NJ系统树上分为两支,其中5.8S r DNA序列基本一致,表现出高度的保守性。2级结构预测结果显示ITS1和ITS2区表现出了较大的种间差异。因此ITS序列可以作为云南丹参、正品药用丹参及其他近缘种的鉴别方法。

Materials from 8 locations areas were selected for the genomie DNA extraction, then ITS sequence was amplified and sequenced. Salvia yunnanensis and Saliva miltiorrhiza were identified and clustered by ITS barcode and its seeondary structure. Sequence alignment were performed by DNAMAN 8.0, the NJ phylogenetie tree was built by MEGA 6.0, the seeondary structure of ITS1 and ITS2 sequences were predicted by RNAstruet 6.0. The results of study show that there is a significant differenee between ITS sequences of S. yunnanensis and S. miltiorrhiza, which is divided into 2 branches in the NJ phylogenetie tree, 5.8S rDNA sequences are basically the same and show a high degree of conservation among them. The seeondary structure indicates that ITS1 and ITS2 regions show a large interspeeifie divergence. Therefore, the ITS sequence eouht provide a basis for the identification of S. yunnanensis , S. miltiorrhiza and other related species.