Development of Multiplexed PCR marker to discriminate between Saposhnikovia divaricata, Peucedaum japonicum and Glehnia littoralis

Abstract

Saposhnikovia divaricata (Umbelliferae) is applied for headaches, vertigo, generalized aching [1]. There are 11 types of Saposhnikovia divaricata in China and Korea and Japan mostly import the materials. In Korea, Peucedaum japonicum and Glehnia littoralis are used for the same purpose, and the three medicines are often confused in prescriptions. The internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA sequence were compared by using fresh leaf and dried root of 31 samples. By comparing the base sequence of the ITS region, we developed molecular markers for the analysis of S. divaricata, P. japonicum and G. littoralis. We designed four primers of BP-F1, BP-F2, BP-R1 and BP-R2. We could develop the multiplex SCAR molecular marker [2, 3] that simultaneously detect products of 364 bp, 191bp, and Saposhnikovia divaricata,494 bp. This multiplex-PCR technique can be used to discriminate these three species in a short period time.