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( Yurry Um ),( Mei Lan Jin ),( Ok Tae Kim ),( Young Chang Kim ),( Seong Cheol Kim ),( Seon Woo Cha ),( Ki Wha Chung ),( Serim Kim ),( Chan Moon Chung ),( Yi Lee ) 한국육종학회 2016 Plant Breeding and Biotechnology Vol.4 No.1
Panax ginseng has been one of the most important herbal medicines used in Eastern Asia. Recently, various molecular markers have been developed to authenticate Panax species, but these markers cannot differentiate the exact varieties or variants of Korean ginseng cultivars. In this study, six cultivars of Korean ginseng (Chunpoong, Yunpoong, Gopoong, Gumpoong, Jakyung, and Hwangsook), P. quinquefolius, and P. notoginseng were differentiated by simple sequence repeat (SSR) marker development. Specific primer sets were designed for the 54 candidate sequences containing SSRs that were predicted. Finally, eight polymorphic SSR loci were developed. DNA fragment analysis was performed using fluorescence-labelled primers for the amplicons. Reproducibility tests were carried out using multiple samples of Korean ginseng cultivars and Panax species. Eight primer sets (PgSSR07, PgSSR08, PgSSR09, PgSSR17, PgSSR37, PgSSR40, PgSSR51, and PgSSR53) showing polymorphism were used for phylogenetic relationship analysis. Consequently, six Korean ginseng cultivars (Chunpoong, Yunpoong, Gopoong, Gumpoong, Jakyung, and Hwangsook), P. quinquefolius, and P. notoginseng could be identified using the combination of SSR markers discovered.
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최홍일,김남훈,김준하,최범순,안인옥,이준수,양태진 고려인삼학회 2011 Journal of Ginseng Research Vol.35 No.4
Little is known about the genetics or genomics of Panax ginseng. In this study, we developed 70 expressed sequence tagderived polymorphic simple sequence repeat markers by trials of 140 primer pairs. All of the 70 markers showed reproducible polymorphism among four Panax species and 19 of them were polymorphic in six P. ginseng cultivars. These markers segregated 1:2:1 manner of Mendelian inheritance in an F2 population of a cross between two P. ginseng cultivars, ‘Yunpoong’ and ‘Chunpoong’,indicating that these are reproducible and inheritable mappable markers. A phylogenetic analysis using the genotype data showed three distinctive groups: a P. ginseng-P. japonicus clade, P. notoginseng and P. quinquefolius, with similarity coefficients of 0.70. P. japonicus was intermingled with P. ginseng cultivars, indicating that both species have similar genetic backgrounds. P. ginseng cultivars were subdivided into three minor groups: an independent cultivar ‘Chunpoong’, a subgroup with three accessions including two cultivars, ‘Gumpoong’ and ‘Yunpoong’ and one landrace ‘Hwangsook’ and another subgroup with two accessions including one cultivar, ‘Gopoong’ and one landrace ‘Jakyung’. Each primer pair produced 1 to 4 bands, indicating that the ginseng genome has a highly replicated paleopolyploid genome structure.
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Yong-Kook Kwon,Myung Suk Ahn,Jong Suk Park,Jang Ryol Liu,Dong Su In,Byung Whan Min,Suk Weon Kim 고려인삼학회 2014 Journal of Ginseng Research Vol.38 No.1
To determine whether Fourier transform (FT)-IR spectral analysis combined with multivariate analysis of whole-cell extracts from ginseng leaves can be applied as a high-throughput discrimination system of cultivation ages and cultivars, a total of total 480 leaf samples belonging to 12 categories corresponding to four different cultivars (Yunpung, Kumpung, Chunpung, and an open-pollinated variety) and three different cultivation ages (1 yr, 2 yr, and 3 yr) were subjected to FT-IR. The spectral data were analyzed by principal component analysis and partial least squares-discriminant analysis. A dendrogram based on hierarchical clustering analysis of the FT-IR spectral data on ginseng leaves showed that leaf samples were initially segregated into three groups in a cultivation age-dependent manner. Then, within the same cultivation age group, leaf samples were clustered into four subgroups in a cultivar-dependent manner. The overall prediction accuracy for discrimination of cultivars and cultivation ages was 94.8% in a cross-validation test. These results clearly show that the FT-IR spectra combined with multivariate analysis from ginseng leaves can be applied as an alternative tool for discriminating of ginseng cultivars and cultivation ages. Therefore, we suggest that this result could be used as a rapid and reliable F1 hybrid seed-screening tool for accelerating the conventional breeding of ginseng.
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Hong-Il Choi,Nam Hoon Kim,Jun Ha Kim,Beom Soon Choi,In-Ok Ahn,Joon-Soo Lee,Tae-Jin Yang 고려인삼학회 2011 Journal of Ginseng Research Vol.35 No.4
Little is known about the genetics or genomics of Panax ginseng. In this study, we developed 70 expressed sequence tag-derived polymorphic simple sequence repeat markers by trials of 140 primer pairs. All of the 70 markers showed reproducible polymorphism among four Panax species and 19 of them were polymorphic in six P ginseng cultivars. These markers segregated 1 :2: I manner of Mendelian inheritance in an F2 population of a cross between two P. ginseng cultivars, "Yunpoong" and "Chunpoong", indicating that these are reproducible and inheritable mappable markers. A phylogenetic analysis using the genotype data showed three distinctive groups: a P. ginseng-P.japonicus clade, P. notoginseng and P. quinquefolius, with similarity coefficients of 0.70. P. japonicus was intermingled with P. ginseng cultivars, indicating that both species have similar genetic backgrounds. P. ginseng cultivars were subdivided into three minor groups: an independent cultivar "Chunpoong", a subgroup with three accessions including two cultivars, "Gumpoong" and "Yunpoong" and one landrace "Hwangsook" and another subgroup with two accessions including one cultivar, "Gopoong" and one landrace "Jakyung". Each primer pair produced I to 4 bands, indicating that the ginseng genome has a highly replicated paleopolyploid genome structure.
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Choi, Hong-Il,Kim, Nam-Hoon,Kim, Jun-Ha,Choi, Beom-Soon,Ahn, In-Ok,Lee, Joon-Soo,Yang, Tae-Jin The Korean Society of Ginseng 2011 Journal of Ginseng Research Vol.35 No.4
Little is known about the genetics or genomics of Panax ginseng. In this study, we developed 70 expressed sequence tagderived polymorphic simple sequence repeat markers by trials of 140 primer pairs. All of the 70 markers showed reproducible polymorphism among four Panax species and 19 of them were polymorphic in six P. ginseng cultivars. These markers segregated 1:2:1 manner of Mendelian inheritance in an $F_2$ population of a cross between two P. ginseng cultivars, 'Yunpoong' and 'Chunpoong', indicating that these are reproducible and inheritable mappable markers. A phylogenetic analysis using the genotype data showed three distinctive groups: a P. ginseng-P. japonicus clade, P. notoginseng and P. quinquefolius, with similarity coefficients of 0.70. P. japonicus was intermingled with P. ginseng cultivars, indicating that both species have similar genetic backgrounds. P. ginseng cultivars were subdivided into three minor groups: an independent cultivar 'Chunpoong', a subgroup with three accessions including two cultivars, 'Gumpoong' and 'Yunpoong' and one landrace 'Hwangsook' and another subgroup with two accessions including one cultivar, 'Gopoong' and one landrace 'Jakyung'. Each primer pair produced 1 to 4 bands, indicating that the ginseng genome has a highly replicated paleopolyploid genome structure.
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Kwon, Yong-Kook,Ahn, Myung Suk,Park, Jong Suk,Liu, Jang Ryol,In, Dong Su,Min, Byung Whan,Kim, Suk Weon The Korean Society of Ginseng 2014 Journal of Ginseng Research Vol.38 No.1
To determine whether Fourier transform (FT)-IR spectral analysis combined with multivariate analysis of whole-cell extracts from ginseng leaves can be applied as a high-throughput discrimination system of cultivation ages and cultivars, a total of total 480 leaf samples belonging to 12 categories corresponding to four different cultivars (Yunpung, Kumpung, Chunpung, and an open-pollinated variety) and three different cultivation ages (1 yr, 2 yr, and 3 yr) were subjected to FT-IR. The spectral data were analyzed by principal component analysis and partial least squares-discriminant analysis. A dendrogram based on hierarchical clustering analysis of the FT-IR spectral data on ginseng leaves showed that leaf samples were initially segregated into three groups in a cultivation age-dependent manner. Then, within the same cultivation age group, leaf samples were clustered into four subgroups in a cultivar-dependent manner. The overall prediction accuracy for discrimination of cultivars and cultivation ages was 94.8% in a cross-validation test. These results clearly show that the FT-IR spectra combined with multivariate analysis from ginseng leaves can be applied as an alternative tool for discriminating of ginseng cultivars and cultivation ages. Therefore, we suggest that this result could be used as a rapid and reliable F1 hybrid seed-screening tool for accelerating the conventional breeding of ginseng.
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Wang, Hongtao,Li, Guisheng,Kwon, Woo-Saeng,Yang, Deok-Chun,Zhu, Jianhua MDPI 2016 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.17 No.6
<P><I>Panax ginseng</I> 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 <I>P. ginseng</I> cultivars, and the EST intron-targeting strategy will provide a potential approach for marker development in species without whole genomic DNA sequence information.</P>
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방경환,조익현,김영창,김장욱,박홍우,신미란,김영배,김옥태,현동윤,김동휘,차선우 韓國藥用作物學會 2012 한국약용작물학회지 Vol.20 No.5
This study was carried out to identify Korean ginseng cultivars using peptide nucleic acid (PNA) microarray. Sixty-seven probes were designed based on nucleotide variation to distinguish Korean ginseng cultivars of Panax ginseng. Among those PNA probes, three (PGB74, PGB110 and PGB130) have been developed to distinguish five Korean ginseng cultivars. Five Korean ginseng cultivars were denoted as barcode numbers depending on their fluorescent signal patterns of each cultivar using three probe sets in the PNA microarray. Five Korean ginseng cultivars, Chunpoong, Yunpoong, Gopoong, Gumpoong and Sunpoong, were simply denoted as '111', '222', '211', '221' and '122', respectively. This is the first report of PNA microarray which provided an objective and reliable method for the authentication of Korean ginseng cultivars. Also, the PNA microarray will be useful for management system and pure guarantee in ginseng seed.
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고려인삼 신품종 ‘천량’ 특이적 DNA 판별 마커 개발
조익현,김영창,김장욱,이승호,임지영,문지영,노봉수,현동윤,김동휘,김기홍,방경환 한국약용작물학회 2014 한국약용작물학회지 Vol.22 No.6
This study describes the efficient method for the discrimination of ‘Cheonryang’ in Panax ginseng Meyerusing a STS primer. A total of 208 STS primers were applied to polymerase chain reaction (PCR) amplification for discriminatingKorean ginseng cultivars. Co-dominant polymorphic band patterns were generated with two primers, MFGp 0019,MFGp 0248, and successful identification of ‘Cheonryang’ was achieved from out of 11 Korean ginseng cultivars. Two differentsizes of DNA band patterns were detected with MFGp 0019 primer. Ten Korean ginseng cultivars shared the samesize of amplified DNAs (389 bp), but ‘Cheonryang’ showed a different size. Thus ‘Cheonryang’ can be efficiently distinguishedfrom the other ten ginseng cultivars by using the MFGp 0019 primer. In the case of MFGp 0248, two different sizesof DNA band patterns were detected in the eleven ginseng cultivars. Same sized amplified DNA bands (307 bp) were shownin five cultivars (Chunpoong, Gopoong, Kumpoong, Cheongsun, Sunhyang) and 254 bp sized DNA bands were identified inthe other 6 cultivars (Yunpoong, Sunpoong, Sunun, Sunone, Cheonryang, K-1). In conclusion, the two STS primers, MFGp0019, and MFGp 0248, provide a rapid and reliable method for the specific identification of ‘Cheonryang’ cultivar from alarge number of samples.
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EST-SSR Marker Sets for Practical Authentication of All Nine Registered Ginseng Cultivars in Korea
Kim, Nam-Hoon,Choi, Hong-Il,Ahn, In-Ok,Yang, Tae-Jin The Korean Society of Ginseng 2012 Journal of Ginseng Research Vol.36 No.3
Panax ginseng has been cultivated for centuries, and nine commercial cultivars have been registered in Korea. However, these nine elite cultivars are grown in less than 10% of ginseng fields, and there is no clear authentication system for each cultivar even though their values are higher than those of local landraces. Here, we have developed 19 microsatellite markers using expressed gene sequences and established an authentication system for all nine cultivars. Five cultivars, 'Chunpoong', 'Sunpoong', 'Gumpoong', 'Sunun', and 'Sunone', can each be identified by one cultivar-unique allele, gm47n-a, gm47n-c, gm104-a, gm184-a (or gm129-a), and gm175-c, respectively. 'Yunpoong' can be identified by the co-appearance of gm47n-b and gm129-c. 'Sunhyang' can be distinguished from the other eight cultivars by the co-appearance of gm47n-b, gm129-b, and gm175-a. The two other cultivars, 'Gopoong' and 'Cheongsun', can be identified by their specific combinations of five marker alleles. This marker set was successfully utilized to identify the cultivars among 70 ginseng individuals and to select true F1 hybrid plants between two cultivars. We further analyzed the homogeneity of each cultivar and phylogenetic relationships among cultivars using these markers. This marker system will be useful to the seed industry and for breeding of ginseng.
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