Development of a single-nucleotide-polymorphism marker for specific authentication of Korean ginseng (Panax ginseng Meyer) new cultivar "G-1"

Abstract

Background: Korean ginseng (Panax ginseng) is a well-known medicinal plant of Oriental medicine that is still in practice today. Until now, a total of 11 Korean ginseng cultivars with unique features to Korean ginseng have been developed based on the pure-line-selection method. Among them, a new cultivar namely G-1 with different agricultural traits related to yield and content of ginsenosides, was developed in 2012.

Methods: The aim of this study was to distinguish the new ginseng cultivar G-1 by identifying the unique single-nucleotide polymorphism (SNP) at its 45S ribosomal DNA and Panax quinquefolius region than other Korean ginseng cultivars using multiplex amplification-refractory mutation system-polymerase chain reaction (ARMS-PCR).

Results: A SNP at position of 45S ribosomal DNA region between G-1, P. quinquefolius, and the other Korean ginseng cultivars was identified. By designing modified allele-specific primers based on this site, we could specifically identified G-1 and P. quinquefolius via multiplex PCR. The unique primer for the SNP yielded an amplicon of size 449 bp in G-1 cultivar and P. quinquefolius. This study presents an effective method for the genetic identification of the G-1 cultivar and P. quinquefolius.

Conclusion: The results from our study shows that this SNP-based approach to identify the G-1 cultivar will be a good way to distinguish accurately the G-1 cultivar and P. quinquefolius from other Korean ginseng cultivars using a SNP at 45S ribosomal DNA region.

Keywords: G-1 cultivar; Panax ginseng; Panax quinquefolius; Single-nucleotide polymorphism; multiplex polymerase chain reaction.

Fig. 1

Comparison of the 45S ribosmal DNA sequences of 11 Korean ginseng cultivars and Panax quinquefolius. The specific primers designed in ribosomal DNA 45S region. KgF: universal primer to Korean ginseng; G-1F: positive primer specific to G-1 cultivar and Panax quinquefolius; AgF: positive primer specific to P. quinquefolius.

Fig. 2

Schematic diagram of the ribosomal 45S region and the positions of the specific primers used for multiplex polymerase chain reaction. G-1, the other Korean ginseng cultivars, and Panax quinquefolius yielded the universal band of 562 bp amplified by primers KgF and 45SR from the ribosomal 45S region.

Fig. 3

Products of multiplex amplification-refractory mutation system–polymerase chain reaction using primers KgF, G-1F, AgF, and 45SR. Lane M: 1,000-bp DNA ladder; lane 1: Chunpoong; lane 2: Yunpoong; lane 3: Gopoong; lane 4: Sunpoong; lane 5: Gumpoong; lane 6: Sunun; lane 7: Chungsun; lane 8: Sunone; lane 9: Sunhyang; lane 10: K-1; lanes 11–13: G-1; lanes 14–16: Panax quinquefolius. G-1 and P. quinquefolius yielded 449 bp and 255 bp for G-1F and AgF, respectively. The experiments were conducted 10 times with a large number of ginseng samples, and the reproducibility of the amplification-refractory-mutation-system results were validated.