Authentication of Panax ginseng from its Adulterants by PCR-RFLP and ARMS

Abstract

As a widely used and expensive herbal medicine, Panax ginseng has many adulterants in the commercial market. PCR-restriction fragment length polymorphism (PCR-RFLP) and amplification refractory mutation system (ARMS) based on 5S rDNA sequence analysis were applied to identify two common adulterants of P. ginseng. The sizes of 5S rRNA gene non-transcribed spacers (NTS) sequences in P. ginseng and its adulterants were determined, ranging from 143 to 424 bp. The PCR product of P. ginseng only could be digested among the tested specimens because of its specific SpeI restriction site found in the 5S rDNA sequence. In addition, P. ginseng was successfully identified from compound medicinal preparations and from the Single-Taste medicines. These results suggest that the methods are able to authenticate P. ginseng.

Key words

Panax ginseng - Araliaceae - authentication - PCR‐RFLP - 5S rDNA NTS - ARMS

References

• 1 Baker W J, Hedderson T A, Dransfield J. Molecular phylogenetics of Calamus (Palmae) and related rattan genera based on 5S rDNA spacer sequence data.  Mol Phyl Evol. 2000;  11 218-231
• 2 Bernard R B, Douglas A J, Grant L B. Ancient differentiation of the H and I haplomes in diploid Hordeum species based on 5S rDNA.  Genome. 2005;  48 610-618
• 3 Li X, Ding X, Chu B, Ding G, Gu S, Qian L, Wang Y, Zhou Q. Molecular authentication of Alisma orientale by PCR-RFLP and ARMS.  Planta Med. 2006;  72 1-4
• 4 Park M J, Kim M K, In J G, Yang D C. Molecular identification of Korean ginseng by amplification refractory mutation system-PCR.  Food Res Int. 2006;  39 568-574
• 5 Cui X M, Lo C K, Yip K L, Dong T T, Tsim K W. Authentication of Panax notoginseng by 5S-rRNA spacer domain and random amplified polymorphic DNA (RAPD) analysis.  Planta Med. 2003;  69 584-586
• 6 Hon C C, Chow Y C, Zeng F Y, Leung F C. Genetic authentication of ginseng and other traditional Chinese medicine.  Acta Pharmacol Sin. 2003;  24 841-846
• 7 Shaw P C, But P. Authentication of Panax species and their adulterants by random-primed polymerase chain reaction.  Planta Med. 1995;  61 466-469
• 8 Xia H Y, You Z H, Shaw P C, Wang J. Application of AFLP technique in authentication of Panax ginseng and Panax quinquefolius. Shaw PC, Chao H, eds. Molecular authentication of Chinese traditional medicine. Shanghai; Fudan Press 2004: 129-141
• 9 Um J Y, Chung H S, Kim M S, Na H J, Kwon H J, Kim J J, Lee K M, Lee S J, Lim J P, Do K R, Hwang W J, Lyu Y S, An N H, Kim H M. Molecular authentication of Panax ginseng species by RAPD analysis and PCR-RFLP.  Biol Pharm Bull. 2001;  24 872-875
• 10 Ngan F, Shaw P, But P, Wang J. Molecular authentication of Panax species.  Phytochemistry. 1999;  50 787-791
• 11 Fukui K, Kamusugi Y, Sakai F. Physical mapping of 5S rDNA loci by direct-cloned biotinylated probes in barley chromosome.  Genome. 1994;  37 105-111
• 12 Thompson J D, Gibson T J, Plewniak F, Jeanmougin F, Higgins D G. The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.  Nucleic Acids Res. 1997;  24 4876-4882