Panax ginseng-specific sequence characterized amplified region (SCAR) marker for testing medicinal products

Abstract

To screen genetic polymorphisms of Panax ginseng, as well as those of Panax quinquefolium and Panax notoginseng, analysis of random amplified polymorphic DNA (RAPD) was performed using 120 random primers. Of the successful amplicons obtained, the Panax ginseng-specific RAPD marker C-12 was cloned into a TA vector and sequenced (GenBank access number KU553472). Based on the sequence analysis results, a pair of primers specific to C-12 was designed. Finally, a SCAR marker-based identification system for Panax ginseng was developed after optimization of the reaction conditions. Using this method, two positive bands were stably observed at 300 bp and 130 bp in 33 batches of Panax ginseng samples tested, while negative results were obtained for another 101 batches of samples, including Panax quinquefolium, Panax notoginseng, adulterants, and other medicinal herbs. Thus, we successfully developed a PCR-based method for rapid and effective identification of Panax ginseng, which can be effectively used for the protection and utilization of germplasm resources and identification of the origins of Panax ginseng samples.

Keywords

Panax ginseng / random amplified polymorphic DNA (RAPD) / sequence characterized amplified regions (SCAR) / molecular identification

References

[1] AtteleA S, WuJ-a, YuanC-su. Ginseng pharmacology: Multiple constituents and multiple actions [J]. Biochemical Pharmacology, 1999, 58(11): 1685-1693

[2] ChangY S, SeoE K, GyllenhaalC, BlockK I. Panax ginseng: a role in cancer therapy? [J]. Integrative Cancer Therapies, 2003, 2(1): 13-33

[3] ChengY, ShenL-h, ZhangJ-tian. Anti-amnestic and anti-aging effects of ginsenoside Rg1 and Rb1 and its mechanism of action [J]. Acta Pharmacologica Sinica, 2005, 26(2): 143-149

[4] ChooM K, ParkE K, HanM J, KimD H. Antiallergic activity of ginseng and its ginsenosides [J]. Planta Medica, 2003, 69(6): 518-522

[5] ShinH R, KimJ Y, YunT K, MorganG, VainioH. The cancer-preventive potential of Panax ginseng: A review of human and experimental evidence [J]. Cancer Causes & Control, 2000, 11(6): 565-576

[6] WangH-w, PengD-c, XieJ-tian. Ginseng leaf-stem: Bioactive constituents and pharmacological functions [J]. Chinese Medicine, 2009, 4(1): 20

[7] HeY-qing. The discuss ginseng market situation and the authenticity of identification [J]. China Practical Medical, 2011, 618226-227

[8] WuX-l, LiuL-l, ZhangZ-z, DengF-f, LiuX-xing. Molecular characterization of Acidithiobacillus ferrooxidans strains isolated from different environments by three PCR-based methods [J]. Journal of Central South University, 2015, 22(4): 455-465

[9] ChenL-j, QiX, WangY-k, ZhangL, GuoZ-f, LinJ-w, SongY-n, ZhongMing. Identification of Schisandra sphenanthera and S. chinensis by random amplified polymorphic DNA sequence characterized applied region] [J]. China Journal of Chinese Materia Medica, 2011, 36(22): 3083-3085

[10] CuiZ-h, LongP, WangY-l, BaiX-r, YuanY, LiM-hui. Application and prospect of DNA molecular markers in the identification of Chinese Medicine [J]. Journal of Chinese Medicinal Materials, 2015, 38(1): 188-192

[11] KiranU, KhanS, MirzaK J, RamM, AbdinM Z. SCAR markers: A potential tool for authentication of herbal drugs [J]. Fitoterapia, 2010, 81(8): 969-976

[12] SunT, TengS-n, KongD-y, SongY, XuJ, LiY-g, WangY, LiM-fu. DNA barcoding used in the identification of ginseng [J]. China Biotechnology, 2013, 33(4): 143-148

[13] ZhouT, XieY, ZhangL-y, WeiS-h, JinY-lei. Study on sequence characterized amplified region (SCAR) markers of polygonum capitatum [J]. China Journal of Chinese Materia Medica, 2013, 38(16): 2577-2580

[14] TanakaH, FukudaN, ShoyamaY. Identification and differentiation of Panax species using ELISA, RAPD and eastern blotting [J]. Phytochemical Analysis, 2006, 17(1): 46-55

[15] JungJ, KimK-h, YangK, BangK-h, YangT-Jin. Practical application of DNA markers for high-throughput authentication of Panax ginseng and Panax quinquefolius from commercial ginseng products [J]. Journal of Ginseng Research, 2014, 38(2): 123-129

[16] PirttiläA M, HirsikorpiM, KämäräinenT, JaakolaL, HohtolaA. DNA isolation methods for medicinal and aromatic plants [J]. Plant Molecular Biology Reporter, 2001, 19(3): 273-273

[17] SolieriL, GiudiciP. Development of a sequencecharacterized amplified region marker-targeted quantitative PCR assay for strain-specific detection of Oenococcus oeni during wine malolactic fermentation [J]. Applied and Environmental Microbiology, 2010, 76(23): 7765-7774

[18] XiaJ-l, ZhangQ, ZhangR-y, PengJ-h, PengA-a, ZhaoX-j, NieZ-y, QiuG-zhou. Isolation and characterization of acidophilic bacterium from Dongxiangshan Mine in Xinjiang Province, China [J]. Journal of Central South University, 2010, 17(1): 50-55