Molecular identification of Schisandra chinensis and its allied species using multiplex PCR based on SNPs

Abstract

Schisandra chinensis (Turcz.) Baill. is a climbing woody vine found over a wide geographical range in northeast Asia. Its red five-flavored fruits have been used as drugs and food. In China and Korea, the dried fruit of S. chinensis has been used for a long time as a traditional medicine. Three species of the family Schisandraceae, S. chinensis, S. repanda, and Kadsura japonica, are distributed mainly in southern Korea. While S. chinensis is broadly distributed, S. repanda and K. japonica are restricted to the southern islands of Korea, especially Jeju. The amplification refractory mutation system (ARMS), one of molecular genotyping technique, is a simple and rapid method for detecting point mutations, restriction fragment length polymorphisms, and small deletions or insertions in a DNA sequence. This PCR-based technique utilizes a primer designed to react only when it anneals to specific target sequences in a DNA sample. In the present study, we analyzed rbcL of the chloroplast gene and ITS from S. chinensis and two related taxa, S. repanda and K. japonica, and developed a specific marker for distinguishing S. chinensis from the related taxa based on SNPs using multiplex PCR. Result showed that two species-specific fragments with 230 bp of rbcL and 278 bp of ITS region were additionally amplified only in all of individuals of S. chinenesis using multiplex PCR method. Here, we describe the details of the protocol for the molecular identification of S. chinensis and other related species. Our results cover five populations of S. chinensis in Korea.