Molecular authentication of Pinelliae Tuber and its common adulterants using RAPD-derived multiplex sequence characterized amplified region (multiplex-SCAR) markers

The identification of authentic plant species is important for the quality control of traditional herbal medicines. The Pinelliae Tuber described as the tuber of only Pinellia ternata in national pharmacopoeia of Korea, China, and Japan. However, the tubers and herbal medicines derived from Pinellia ternata are similar to those derived from P. tripartita, P. pedatisecta, and Typhonium flagelliforme, and the correct identification of species is very difficult using conventional methods. Therefore, this study was carried out to develop DNA based markers for the identification authentic Pinelliae Tuber and its common adulterants in species levels. To develop a reliable method to discriminate Pinelliae Tuber and its closely related three common adulterants, we introduced sequence characterized amplification region (SCAR) markers using randomly amplified polymorphic DNA (RAPD)-based SCAR methods. Several distinct SCAR markers were developed that amplified unique single DNA fragments in P. ternata, P. tripartita, P. pedatisecta, and T. flagelliforme based on the verification of species-specific RAPD amplicons. Furthermore, a useful molecular marker was established for multiplex PCR, enabling the four species to be distinguished concurrently in a single PCR reaction. These genetic markers enable the efficient and rapid discrimination of authentic Pinelliae Tuber, the herbal medicine derived from the tuber of P. ternata, from closely related adulterant species and will be useful for the standardization of this herbal medicine.