Molecular Identification of Salvia miltiorrhiza Bge. and Its Adulterants Based on ITS2 Sequences

摘要

[Objectives]The study was conducted to investigate the molecular identification of Salvia miltiorrhiza Bge. and its adulterants by DNA barcoding andspecific primer PCR. [Methods]With ITS2 sequenceas DNA barcode,the materials were amplified by PCR and sequenced,and the NJ phylogenetic tree was constructed. The secondary structure of ITS2 was predicted by database and its website established by Koetschan et al.,and the self-designed primers were used to carry out specific primer PCR identification. [Results]ITS2 sequence length was around 470 bp. The results of cluster analysis showed that S. miltiorrhiza Bge. and its adulterants were clustered on different branches and showed monophyly. The comparison of secondary structure showed that S. miltiorrhiza Bge. had little differences from S. przewalskii,while there were significant differences from A. lappa in the number,size and location of stem-loop and the rotation angle of the spiral arm from the central ring. The specific primers could distinguish the S. miltiorrhiza Bge. and its counterfeits by PCR technique. [Conclusions] DNA barcoding and specific primer PCR are effective in distinguishing S. miltiorrhiza Bge. and its adulterants,and it has an important application foreground in the identification of Chinese herbal medicines.