Molecular discrimination of Panax ginseng cultivar K-1 using pathogenesis-related protein 5 gene.

BACKGROUND: The mixed-cultivation of different Panax ginseng cultivars can cause adverse effects on stability of yield and quality. K-1 is a superior cultivar with good root shape and stronger disease resistance. DNA markers mined from functional genes are clearly desirable for K-1, as they may associate with major traits and can be used for marker-assisted selection to maintain the high quality of Korean ginseng. METHODS: Five genes encoding pathogenesis-related (PR) proteins of P. ginseng were amplified and compared for polymorphism mining. Primary, secondary, and tertiary structures of PR5 protein were analyzed by ExPASy-ProtParam, PSSpred, and I-TASSER methods, respectively. A coding single nucleotide polymorphism (SNP)-based specific primer was designed for K-1 by introducing a destabilizing mismatch within the 3' end. Allele-specific polymerase chain reaction (PCR) and real-time allele-specific PCR assays were conducted for molecular discrimination of K-1 from other cultivars and landraces. RESULTS: A coding SNP leading to the modification of amino acid residue from aspartic acid to asparagine was exploited in PR5 gene of K-1 cultivar. Bioinformatics analysis showed that the modification of amino acid residue changed the secondary and tertiary structures of the PR5 protein. Primer KSR was designed for specific discrimination of K-1 from other ginseng cultivars and landraces. The developed real-time allele-specific PCR assay enabled easier automation and accurate genotyping of K-1 from a large number of ginseng samples. CONCLUSION: The SNP marker and the developed real-time allele-specific PCR assay will be useful not only for marker-assisted selection of K-1 cultivar but also for quality control in breeding and seed programs of P. ginseng.

Molecular differentiation of Russian wild ginseng using mitochondrial nad7 intron 3 region.

BACKGROUND: Cultivated ginseng is often introduced as a substitute and adulterant of Russian wild ginseng due to its lower cost or misidentification caused by similarity in appearance with wild ginseng. The aim of this study is to develop a simple and reliable method to differentiate Russian wild ginseng from cultivated ginseng. METHODS: The mitochondrial NADH dehydrogenase subunit 7 (nad7) intron 3 regions of Russian wild ginseng and Chinese cultivated ginseng were analyzed. Based on the multiple sequence alignment result, a specific primer for Russian wild ginseng was designed by introducing additional mismatch and allele-specific polymerase chain reaction (PCR) was performed for identification of wild ginseng. Real-time allele-specific PCR with endpoint analysis was used for validation of the developed Russian wild ginseng single nucleotide polymorphism (SNP) marker. RESULTS: An SNP site specific to Russian wild ginseng was exploited by multiple alignments of mitochondrial nad7 intron 3 regions of different ginseng samples. With the SNP-based specific primer, Russian wild ginseng was successfully discriminated from Chinese and Korean cultivated ginseng samples by allele-specific PCR. The reliability and specificity of the SNP marker was validated by checking 20 individuals of Russian wild ginseng samples with real-time allele-specific PCR assay. CONCLUSION: An effective DNA method for molecular discrimination of Russian wild ginseng from Chinese and Korean cultivated ginseng was developed. The established real-time allele-specific PCR was simple and reliable, and the present method should be a crucial complement of chemical analysis for authentication of Russian wild ginseng.

Cytological analysis of ginseng carpel development.

Panax ginseng Meyer, commonly known as ginseng, is considered one of the most important herbs with pharmaceutical values due to the presence of ginsenosides and is cultivated for its highly valued root for medicinal purposes. Recently, it has been recognized that ginseng fruit contains high contents of triterpene such as ginsenoside Re as pharmaceutical compounds. However, it is unclear how carpel, the female reproductive tissue of flowers, is formed during the three-year-old growth before fruit is formed in ginseng plants. Here, we report P. ginseng carpel development at the cytological level, starting from the initial stage of ovule development to seed development. The carpel of P. ginseng is composed of two free stigmas, two free styles, and one epigynous bilocular ovary containing one ovule in each locule. Based on our cytological study, we propose that the female reproductive development in P. ginseng can be classified into seven stages: early phase of ovule development, megasporogenesis, megagametogenesis, pre-fertilization, fertilization, post-fertilization, and seed development. We also describe the correlation of the female and male gametophyte development and compare morphological differences in carpel development between ginseng and other higher plants. One unique feature for ginseng seed development is that it takes 40 days for the embryo to develop to the early torpedo stage and that the embryo is small relative to the seed size, which could be a feature of taxonomic importance. This study will provide an integral tool for the study of the reproductive development and breeding of P. ginseng.

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

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.

Discrimination of Korean ginseng (Panax ginseng Meyer) cultivar Chunpoong and American ginseng (Panax quinquefolius) using the auxin repressed protein gene.

BACKGROUND: Korean ginseng (Panax ginseng) is one of the most important medicinal plants in the Orient. Among nine cultivars of P. ginseng, Chunpoong commands a much greater market value and has been planted widely in Korea. Chunpoong has superior quality "Chunsam" (1st grade ginseng) when made into red ginseng. METHODS: A rapid and reliable method for discriminating the Chunpoong cultivar was developed by exploiting a single nucleotide polymorphism (SNP) in the auxin repressed protein gene of nine Korean ginseng cultivars using specific primers. RESULTS: An SNP was detected between Chunpoong and other cultivars, and modified allele-specific primers were designed from this SNP site to specifically identify the Chunpoong cultivar and P. quinquefolius via multiplex polymerase chain reaction (PCR). CONCLUSION: These results suggest that great impact to prevent authentication of precise Chunpoong and other cultivars using the auxin repressed protein gene. We therefore present an effective method for the authentication of the Chunpoong cultivar of P. ginseng and P. quinquefolius.

Development of EST Intron-Targeting SNP Markers for Panax ginseng and Their Application to Cultivar Authentication.

Panax ginseng is one of the most valuable medicinal plants in the Orient. The low level of genetic variation has limited the application of molecular markers for cultivar authentication and marker-assisted selection in cultivated ginseng. To exploit DNA polymorphism within ginseng cultivars, ginseng expressed sequence tags (ESTs) were searched against the potential intron polymorphism (PIP) database to predict the positions of introns. Intron-flanking primers were then designed in conserved exon regions and used to amplify across the more variable introns. Sequencing results showed that single nucleotide polymorphisms (SNPs), as well as indels, were detected in four EST-derived introns, and SNP markers specific to "Gopoong" and "K-1" were first reported in this study. Based on cultivar-specific SNP sites, allele-specific polymerase chain reaction (PCR) was conducted and proved to be effective for the authentication of ginseng cultivars. Additionally, the combination of a simple NaOH-Tris DNA isolation method and real-time allele-specific PCR assay enabled the high throughput selection of cultivars from ginseng fields. The established real-time allele-specific PCR assay should be applied to molecular authentication and marker assisted selection of P. ginseng cultivars, and the EST intron-targeting strategy will provide a potential approach for marker development in species without whole genomic DNA sequence information.

Development of interspecies hybrids to increase ginseng biomass and ginsenoside yield.

KEY MESSAGE: Interspecific hybrids between Panax ginseng and P. quinquefolius results in hybrid vigor and higher ginsenoside contents. Ginseng is one of the most important herbs with valued pharmaceutical effects contributing mainly by the presence of bioactive ginsenosides in the roots. However, ginseng industry is impeded largely by its biological properties, because ginseng plants are slow-growing perennial herbs with lower yield. To increase the ginseng yield and amounts of ginsenosides, we developed an effective ginseng production system using the F(1) progenies obtained from the interspecific reciprocal cross between two Panax species: P. ginseng and P. quinquefolius. Although hybrid plants show reduced male fertility, F(1) hybrids with the maternal origin either from P. ginseng or P. quinquefolius displayed heterosis; they had larger roots and higher contents of ginsenosides as compared with non-hybrid parental lines. Remarkably, the F(1) hybrids with the maternal origin of P. quinquefolius had much higher ginsenoside contents, especially ginsenoside Re and Rb1, than those with the maternal origin of P. ginseng. Additionally, non-targeted metabolomic profiling revealed a clear increase of a large number of primary and secondary metabolites including fatty acids, amino acids and ginsenosides in hybrid plants. To effectively identify the F(1) hybrids for the large-scale cultivation, we successfully developed a molecular marker detection system for discriminating F(1) reciprocal hybrids. In summary, this work provided a practical system for reciprocal hybrid ginseng production, which would facilitate the ginseng production in the future.

Cytological characterization of anther development in Panax ginseng Meyer.

Ginseng (Panax ginseng), a valued medicinal herb, is a slow-growing plant that flowers after 3 years of growth with the formation of a solitary terminal umbel inflorescence. However, little is known about cytological events during ginseng reproduction, such as the development of the male organ, the stamen. To better understand the mechanism controlling ginseng male reproductive development, here, we investigated the inflorescence and flower structure of ginseng. Moreover, we performed cytological analysis of anther morphogenesis and showed the common and specialized cytological events including the formation of four concentric cell layers surrounding male reproductive cells followed by subsequent cell differentiation and degeneration of tapetal cells, as well as the formation of mature pollen grains via meiosis and mitosis during ginseng anther development. Particularly, our transverse section and microscopic observations showed that the ginseng tapetal layer exhibits obvious nonsynchronous cell division evidenced by the observation of one or two tapetal layers frequently observed in one anther lobe, suggesting the unique control of cell division. To facilitate the future study on ginseng male reproduction, we grouped the anther development into 10 developmental stages according to the characterized cytological events.

Characteristics of Korean ginseng varieties of Gumpoong, Sunun, Sunpoong, Sunone, Cheongsun, and Sunhyang.

BACKGROUND: Ginseng (Panax ginseng Meyer) is an important medicinal herbs in Asia. However, ginseng varieties are less developed. METHOD: To developed ginseng varieties, a pure line selection method was applied in this study. RESULTS: Gumpoong was testing of 4-yr-old specimens in 2002, the proportions of the below-ground roots that were rusty colored for Gumpoong was 1.29 in Daejeon and 1.45 in Eumseong, whereas the proportions for its yellow berry variant were 2.60 and 2.45 in the two regions, respectively. Thus the Gumpoong was resistant to root rust. Sunpoong has a high yielding property. Its average root weight is 70.6 g for 6-yr-old roots. Its yield is 2.9 kg/1.62m(2) and the rate of heaven- and earth-grade product is 20.9%, which is very high compared to 9.4% for Yunpoong. Sunone is resistance to root rot and the survival rate of 4-yr-old roots was 44.4% in 1997, whereas that of the violet-stem variant landrace was 21.7%. Sunhyang has content of arginyl-fructosyl-glucose (AFG), which produces the unique scent of red ginseng, is 95.1 µmol/g and greater than the 30.8 µmol/g of Chunpoong in 6-yr-old plants. Sunun and Cheongsun are being nurtured to protect genetic resources. CONCLUSION: Developed ginsneg varieties will be used as the basis for the protection of genetic resources and breeding.

Investigation of ginsenosides in different tissues after elicitor treatment in Panax ginseng.

BACKGROUND: The effect of methyl jasmonate (MJ) on ginsenoside production in different organs of ginseng (Panax ginseng Meyer) was evaluated after the whole plant was dipped in an MJ-containing solution. MJ can induce the production of antioxidant defense genes and secondary metabolites in plants. In ginseng, MJ treatment in adventitious root resulted in the increase of dammarenediol synthase expression but a decrease of cycloartenol synthase expression, thereby enhancing ginsenoside biosynthesis. Although a previous study focused on the application of MJ to affect ginsenoside production in adventitious roots, we conducted our research on entire plants by evaluating the effect of exogenous MJ on ginsenoside production with the aim of obtaining new approaches to study ginsenoside biosynthesis response to MJ in vivo. METHODS: Different parts of MJ-treated ginseng plants were analyzed for ginsenoside contents (fine root, root body, epidermis, rhizome, stem, and leaf) by high-performance liquid chromatography. RESULTS: The total ginsenoside content of the ginseng root significantly increased after 2 d of MJ treatment compared with the control not subjected to MJ. Our results revealed that MJ treatment enhances ginsenoside production not in the epidermis but in the stele of the ginseng root, implying transportation of ginsenosides from the root vasculature to the epidermis. Application of MJ enhanced protopanaxadiol (PPD)-type ginsenosides, whereas chilling treatment induced protopanaxatriol (PPT)-type ginsenosides. CONCLUSION: These findings indicate that the production of PPD-type and PPT-type ginsenosides is differently affected by abiotic and biotic stresses in the ginseng plant, and they might play different defense mechanism roles.

Ginsenoside profiles and related gene expression during foliation in Panax ginseng Meyer.

Panax ginseng is one of the most important medicinal plants in Asia. Triterpene saponins, known as ginsenosides, are the major pharmacological compounds in P. ginseng. The present study was conducted to evaluate the changes in ginsenoside composition according to the foliation stage of P. ginseng cultured in a hydroponic system. Among the three tested growth stages (closed, intermediate, and opened), the highest amount of total ginsenoside in the main and fine roots was in the intermediate stage. In the leaves, the highest amount of total ginsenoside was in the opened stage. The total ginsenoside content of the ginseng leaf was markedly increased in the transition from the closed to intermediate stage, and increased more slowly from the intermediate to opened leaf stage, suggesting active biosynthesis of ginsenosides in the leaf. Conversely, the total ginsenoside content of the main and fine roots decreased from the intermediate to opened leaf stage. This suggests movement of ginsenosides during foliation from the root to the leaf, or vice versa. The difference in the composition of ginsenosides between the leaf and root in each stage of foliation suggests that the ginsenoside profile is affected by foliation stage, and this profile differs in each organ of the plant. These results suggest that protopanaxadiol- and protopanaxatriol (PPT)-type ginsenosides are produced according to growth stage to meet different needs in the growth and defense of ginseng. The higher content of PPT-type ginsenosides in leaves could be related to the positive correlation between light and PPT-type ginsenosides.

Molecular characterization of two glutathione peroxidase genes of Panax ginseng and their expression analysis against environmental stresses.

Glutathione peroxidases (GPXs) are a group of enzymes that protect cells against oxidative damage generated by reactive oxygen species (ROS). GPX catalyzes the reduction of hydrogen peroxide (H2O2) or organic hydroperoxides to water or alcohols by reduced glutathione. The presence of GPXs in plants has been reported by several groups, but the roles of individual members of this family in a single plant species have not been studied. Two GPX cDNAs were isolated and characterized from the embryogenic callus of Panax ginseng. The two cDNAs had an open reading frame (ORF) of 723 and 681bp with a deduced amino acid sequence of 240 and 226 residues, respectively. The calculated molecular mass of the matured proteins are approximately 26.4kDa or 25.7kDa with a predicated isoelectric point of 9.16 or 6.11, respectively. The two PgGPXs were elevated strongly by salt stress and chilling stress in a ginseng seedling. In addition, the two PgGPXs showed different responses against biotic stress. The positive responses of PgGPX to the environmental stimuli suggested that ginseng GPX may help to protect against environmental stresses.

A simple and rapid technique for the authentication of the ginseng cultivar, Yunpoong, using an SNP marker in a large sample of ginseng leaves.

Yunpoong is an important Korean ginseng (Panax ginseng C. A. Meyer) cultivar, but no molecular marker has been available to identify Yunpoong from other cultivars. In this study, we developed a single nucleotide polymorphism (SNP) marker for Yunpoong based on analysis of expressed sequence tags (ESTs) in an exon region of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene. This SNP marker had high specificity to authenticate Yunpoong in twelve different main ginseng cultivars. For application of the molecular marker, a rapid identification method was established based on the NaOH-Tris method and real-time polymerase chain reaction (PCR) in order to ensure more efficiency in the cultivar selection. The biggest feature of the NaOH-Tris method was that it made the extraction of DNA very simple and rapid in young leaf tissues. We only spent 1 min to extract DNA and directly used it to do PCR. In this report, the conventional DNA extraction method was used to develop molecular marker process, and the NaOH-Tris method was applied in screening large numbers of cultivars. Moreover, the greatest advantage of the real-time PCR compared with traditional PCR, is time saving and high efficiency. Thus, this strategy provides a rapid and reliable method for the specific identification of Yunpoong in a large number of samples.

Development of a multiplex amplification refractory mutation system for simultaneous authentication of Korean ginseng cultivars "Gumpoong" and "Chungsun".

BACKGROUND AND AIMS. Molecular authentication of Korean ginseng cultivars was investigated using the mitochondrial nicotinamide adenine dinucleotide (NADH) dehydrogenase subunit 7 (nad7) intron 3 region. MATERIALS AND METHODS. A mutation site specific to Panax ginseng "Gumpoong" and "Chungsun" cultivars was detected within the sequence data. Based on this mutation site and the "Gumpoong"-specific single nucleotide polymorphism site reported in 26S rDNA, two modified allele-specific primer pairs were designed and a multiplex amplification refractory mutation system (MARMS) was applied to identify "Gumpoong" and "Chungsun." RESULTS. The results showed that "Gumpoong" and "Chungsun" can be clearly discriminated from the other Korean ginseng cultivars by simultaneously identifying the haplotype of "Gumpoong" and the specific allele of "Chungsun" by applying the MARMS. CONCLUSION. This study, therefore, provides a simple and reliable method for simultaneous authentication of "Gumpoong" and "Chungsun" cultivars.

Molecular authentication of Panax ginseng and ginseng products using robust SNP markers in ribosomal external transcribed spacer region.

Panax ginseng and Panax quinquefolius are the most widely used Panax species, but they are known to have different properties and medicinal values. The aim of this study is to develop a robust and accurate DNA marker for identifying P. ginseng and the origins of ginseng products. Two single nucleotide polymorphism (SNP) sites specific to P. ginseng were exploited from nuclear ribosomal external transcribed spacer (ETS) region. Based on the SNP sites, two specific primers were designed for P. ginseng and P. quinquefolius respectively. P. ginseng can be easily discriminated from P. quinquefolius by amplifying the two specific alleles using multiplex allele-specific PCR. Favorable results can also be obtained from commercial ginseng products. The established method is highly sensitive and can detect 1% of intentional adulteration of P. quinquefolius into P. ginseng down to the 0.1ng level of total DNA. Therefore this study provides a reliable and simple DNA method for authentication of the origins and purities of ginseng products.

Development of molecular markers for the determination of the new cultivar 'Chunpoong' in Panax ginseng C. A. Meyer associated with a major latex-like protein gene.

Chunpoong is one of the most valuable cultivars of Panax ginseng C. A. MEYER, and is widely grown in Korea and China. Insertion/deletion (InDel) markers and single nucleotide polymorphism (SNP) markers are useful tools for marker-assisted selections. The SNP marker for determinate Chunpoong was previously developed from the nad7 gene of mtDNA by Wang et al. (2009) but was effective only on a limited range of cultivars. In this study, we studied the reasons for this limited application and developed new useful markers for application in Chunpoong-breeding programs. The new markers of InDel and SNP were designed in the major latex-like protein (MLP-like) gene which was highly expressed in 4-year-old Chunpoong expressed sequence tags (ESTs). To validate the marker in polymerase chain reaction (PCR), we used an InDel marker for identification of Chunpoong in the 70 Panax samples based on a double-blind test, and the success rate was 100%. For rapid and reliable assay of Chunpoong in numerous samples, we utilized an EvaGreen dye and melting curve method on real-time PCR. Furthermore, we designed an SNP marker that depended on the InDel region for more efficient detection of Chunpoong in real-time PCR. Compared with PCR-based assays, our Chunpoong SNP marker and real-time PCR assay offers a significant savings of time and labor in the analysis of large numbers of Chunpoong samples.

Polygalacturonase inhibiting protein: isolation, developmental regulation and pathogen related expression in Panax ginseng C.A. Meyer.

Polygalacturonase inhibiting proteins (PGIPs) are the major defense proteins which play an important role in resistance to infection of pathogens. A putative novel gene encoding PGIP was isolated from Panax ginseng C.A. Meyer, which shows 70.3 and 68.4% homology with chick pea and Arabidopsis PGIPs. The RACE PCR was preformed to isolate the full-length PGIP cDNA from Panax ginseng. Sequence analysis revealed that the cDNA of PgPGIP is of 1,275 bp in length and that it's containing ORF encodes for a polypeptide of 366 amino acids. Domain analysis revealed that the deduced amino acid sequences of PgPGIP have a typical PGIP topology. The transcription level of PgPGIP was up-regulated in ginseng in response to wounding and infection with phytopathogenic fungi i.e., Rhizoctonia solani, Fusarium oxysporum, Phythium ultimum, Botrytis cinerea, Colletotrichum gloeosporioides and Cylindrocarpon destructans, which causes drastic damage in ginseng plants. The constitutive PgPGIP expression of 4 years old plant, showed elevated transcript level, especially roots, showed maximum then buds, stems and leaves, indicating that the gene is developmentally regulated. The crude PGIP extracts derived from the fungal infected plants directly reduces the aggressive potential of PGs from diverse group of fungi. Like other PGIPs, PgPGIP also possess board spectrum of inhibitory activity. Thus, the presence of PgPGIP gene and their active role in defense mechanism was proved. The structural model of PgPGIP was predicted based on the alignment generated by EBI-Align, the program "MOODELLER" and the predicted structure showed 10 ß-strands and 10 α-helixes region.

A simplified method for identifying the Panax ginseng cultivar Gumpoong based on 26S rDNA.

Molecular identification of PANAX GINSENG (Korean ginseng) cultivars is very important for their management. We have developed a simple and reliable method for specific identification of the superior cultivar "Gumpoong." The 26S rDNA was targeted for our molecular analysis, and a single-nucleotide polymorphism (SNP) was identified between Gumpoong and the other cultivars within the sequence data. From this SNP site, a modified allele-specific primer and a novel primer set have been developed to identify the Gumpoong cultivar via multiplex PCR. The established multiplex PCR method was determined to be effective, and the SNP marker showed high specificity to Gumpoong. Therefore, the described method may serve as a useful tool for rapid selection of Gumpoong cultivar.

A PCR-based SNP marker for specific authentication of Korean ginseng (panax ginseng) cultivar "Chunpoong".

Korean ginseng (Panax ginseng) has been developed as a horticultural crop due to the increasing demand in the world market. "Chunpoong" is an economically important cultivar with superior quality and high yield among nine cultivars of Korean ginseng. The aim of this work was to develop a simple technique for specific authentication of Chunpoong using DNA method. Molecular authentication of Chunpoong was investigated using DNA sequences of mitochondrial cytochrome oxidase subunit 2 (cox2) intron I and intron II regions. A single nucleotide polymorphism (SNP) specific to Chunpoong was detected and amplification refractory mutation system (ARMS)-PCR method was applied to specific identification of Chunpoong based on the SNP site. Ginseng samples collected from other locations were used to validate the SNP marker and the established method was determined to be effective. Thus, this work provides a rapid and reliable method for the specific identification of Chunpoong cultivar.

Molecular identification of the Korean ginseng cultivar "Chunpoong" using the mitochondrial nad7 intron 4 region.

BACKGROUND AND AIMS: Korean ginseng (Panax ginseng) is one of the most important medicinal plants in the Orient. Among the nine cultivars of Korean ginseng, "Chunpoong" commands a much greater market value and has been planted widely. MATERIALS AND METHODS: A rapid and reliable method for discriminating the Chunpoong cultivar was developed by exploiting a single nucleotide polymorphism (SNP) in the mitochondrial nad7 intron 4 region of nine Korean ginseng cultivars using universal primers. RESULTS: A SNP was detected between Chunpoong and other cultivars, and modified allele-specific primers were designed from this SNP site to specifically identify Chunpoong cultivar via multiplex PCR. CONCLUSION: We therefore present an effective method for the genetic identification of the Chunpoong cultivar of ginseng.