Development of SCAR Markers for the Discrimination of Three Species of Medicinal Plants, Angelica decursiva (Peucedanum decursivum), Peucedanum praeruptorum and Anthricus sylvestris, Based on the Internal Transcribed Spacer (ITS) Sequence and Random Amplified Polymorphic DNA (RAPD)

摘要

Angelicae Decursivae Radix ('Jeonho' in Korean) is prescribed as the root of Angelica decursiva (=Peucedanum decursivum) and Peucedanum praeruptorum in Korean pharmacopoeia. However, Anthricus sylvestris has been usually distributed on the market because it is identical to the Korean plant name 'Jeonho'. Furthermore, due to the morphological similarity of the aerial parts and herbal medicines, the correct identification of these roots is difficult. Therefore, to develop a reliable method for discriminating among A. decursiva (= P decursivum), P. praeruptorum and A. sylvestris, we applied the tools of molecular genetics, such as the analysis of ribosomal DNA internal transcribed spacer (rDNA-ITS) and random amplified polymorphic DNA (RAPD). In the comparison of rDNA-ITS sequences, we found a specific primer region for the identification of A. sylvestris among three varieties of the herb that produced a 273 bp strand of DNA specific to A. sylvestris. As the result of RAPID analysis, we developed one sequence characterized amplified region (SCAR) marker for A. decursiva and R praeruptorum that amplified a 363 bp DNA fragment specific to both A. decursiva and P praeruptorum and two markers for P praeruptorum that amplified 145 bp and 305 bp DNA fragments specific to P praeruptorum. Furthermore, we established the SCAR markers for the simultaneous discrimination of the three species by applying a multiplex-polymerase chain reaction (PCR) with a combination of primers. This method of discrimination would be useful in preventing the distribution of adulterates because it can identify each herb and distinguish it from inauthentic substitutions.