Molecular Authentication and Genetic Polymorphism of Korean Ginseng (Panax ginseng C. A. Meyer) by Inter-Simple Sequence Repeats (ISSRs) Markers

Bang, Kyong-Hwan,Lee, Sung-Woo,Hyun, Dong-Yun,Cho, Joon-Hyeong,Cha, Seon-Woo,Seong, Nak-Sul,Huh, Man-Kyu Korean Society of Life Science 2004 생명과학회지 Vol.14 No.3

ISSR마크를 사용하여 고려 인삼의 품종 및 계통간 분자적 인증과 유전적 다형현상을 조사하였다. 56개의 ISSR 프라이머 중 5개가 일곱 품종 및 계통간 명확하고 재현성이 높은 DNA분절을 나타내는 최적 프라이머로 선택되었다. 전체 43밴드는 250 bp - 1,700 bp의 분자량을 가지며 프라이머당 8.6개의 밴드를 나타내었다. 고려 인삼에서 다형현상 정도는 20.9%였다. 특히 천풍 품종이 가장 높은 다형현상을 나타낸 반면 다른 품종은 거의 다형현상을 나타내지 않았다. 결론적으로 DNA수준에서 ISSR마크로 천풍이 다른 고려 인삼의 품종 및 계통인 연풍, 황숙종, 자경종과 구분에 이용될 수 있음이 판명되었다. Molecular authentication and genetic polymorphism of Korean ginseng cultivars and accessions were investigated using ISSR (inter-simple sequence repeat amplification) markers. Five primers among 56 produced clear and reproducible DNA fragments among seven cultivars and accessions. A total of 43 bands ranging from 250 bp to 1,700 bp from five primers were scored. Average number of bands per primer was 8.6 and only nine bands were polymorphic across the six Panax ginseng from Korea. Especially Chunpoong cultivar exhibited the highest level of polymorphism, whereas other accessions did not showed almost any polymorphism. Consequently, these ISSR markers will be available to differentiate Chunpoong cultivar from other major Korean ginseng cultivars and accessions, such as Yunpoong, Hwangsukjong and Jakyungjong, at the DNA level.

Discrimination of Species Specific DNA Markers Using RAPD and AFLP Analysis between Atractylodes japonica Koidz. and Atractylodes macrocephala Koidz.

Kyong-Hwan Bang,Chun-Geon Park,Dong-Chun Jin,Hong-Sig Kim,Hee-Woon Park,Chung-Heon Park,Nak-Sul Seong 韓國藥用作物學會 2003 한국약용작물학회지 Vol.11 No.4

To identify the variation of the RAPD patterns between two Atractylodes species, 52 kinds of random primers were applied to each eight of A japonica and A. macrocephala genomic DNA. Ten primers of 52 primers could be used to discriminate between the species and 18 polymorphisms among 67 scored DNA fragments (18 fragments are specific for A. japonica and A. macrocephala) were generated using these primers, 26.9% of which were polymorphic. RAPD data from the 10 primers was used for cluster analysis. The cluster analysis of RAPD markers showed that the two groups are genetically distinct. On the other hand, to identify the variation of the AFLP patterns and select the species specific AFLP markers, eight combinations of EcoRI/MseI primers were applied to the bulked A. japonica and A. macrocephala genomic DNA. Consequently, three combinations of EcoRI/MseI primers (EcoRI /Mse I ; AAC/CTA, AAC/CAA, AAG/CTA) used in this study revealed 176 reliable AFLP markers, 42.0% of which were polymorphic. 74 polymorphisms out of 176 scored DNA fragments were enough to clearly discriminate between two Atractylodes species.

Physical Mapping of the Species-Specific RAPD Fragments and Telomere Sequence Repeats between Atractylodes japonica and Atractylodes macrocephala

Bang, Kyong Hwan,Koo, Dal Hoe,Huh, Man Kyu,Seong, Nak Sul,Bang, Jae Wook 한국유전학회 2004 Genes & Genomics Vol.26 No.1

This study examined the chromosomal characteristics of Atractylodes japonical and A. macrocephala. Cytogenetic analysis based on the physical mapping of the species-specific RAPD fragments and the telomere sequence repeats was performed using fluorescence in situ hybridization (FISH). Physical mapping using FISH showed that the signals for the species-specific RAPD fragments, AjR1 and AmR1, were distributed over the entire chromosome and exhibited species-specific characteristics. Therefore, these two markers can be used for the chromosomal identification of A. japonica and A. macrocephala. Strong signals for the telomere sequences repeats were detected at the ends of each chromatid in both plants. However, one pair of chromosomes (chromosomes 2) in A. macrocephala did not show this telomeric signal. In conclusion, telomere sequence repeats can be used as an efficient chromosomal marker for discriminating between A. japonica and A. macrocephala.

Construction of Genomic DNA Library of Korean Ginseng (<i>Panax ginseng</i> C. A. M<small>EYER</small>) and Development of Sequence-Tagged Sites

Bang, Kyong-Hwan,Lee, Jei-Wan,Kim, Young-Chang,Kim, Dong-Hwi,Lee, Eung-Ho,Jeung, Ji-Ung Pharmaceutical Society of Japan 2010 Biological & pharmaceutical bulletin Vol.33 No.9

<P>This study describes an efficient approach for developing sequence tagged sites (STS) for <I>Panax ginseng</I> C.A. M<SMALL>EYER</SMALL>, and their applications for line discrimination. By using the methylation filtering (MF) technique, a genomic library was constructed, in which clone inserts were derived from the hypomethylated regions of ginseng genome. A methylation unfiltered genomic library was also constructed and the clone inserts were compared to those from the MF library in terms of sequence characteristics. Sequence analysis revealed that MF efficiently enriched the protein coding region of <I>P. ginseng</I>, for which the repetitive DNA appeared to be as little as 2.5 fold lower than clones in the unfiltered library, and also indicated that the <I>P. ginseng</I> genome may contain a large fraction of methylated repetitive DNA elements. A total of 99 and 100 highly stringent STS primer sets were designed from the filtered and unfiltered library, respectively. Amplification products were tested for latent polymorphism across six cultivars of <I>P. ginseng</I> and other 2 <I>Panax</I> species using six endonucleases recognizing four-bases. STS primer sets described here will be useful for marker-assisted selection, genome mapping and line discrimination of <I>P. ginseng</I> or its cultivars from other <I>Panax</I> species.</P>

Comparative Cytogenetic Characteristics and Physical Mapping of the 17S and 5S Ribosomal DNAs between Atractylodes japonica Koidz. and Atractylodes macrocephala Koidz.

Bang, Kyong-Hwan,Koo, Dal-Hoe,Kim, Hong-Sig,Song, Beom-Heon,Cho, Yong-Gu,Cho, Joon-Hyeong,Bang, Jae-Wook The Korean Society of Medicinal Crop Science 2003 韓國藥用作物學會誌 Vol.11 No.4

This study was carried out to compare chromosomal characteristics between Atractylodes japonica and A macrocephala. Cytogenetic analysis was conducted based on karyotype analysis and physical mapping using fluorescence in situ hybridization. As a result of karyotype analysis by feulgen staining, somatic chromosome numbers of A. japonica and A. macrocephala were 2n=24. The length. of the mitotic metaphase chromosomes of A. japonica ranged from $0.70\;to\;1.60{\mu}m$ with a total length. of $12.11{\mu}m$ and the homologous chromosome complement comprised six metacentrics, five submetacentrics and one subtelocentrics. On the other hand, the length of the mitotic metaphase chromosomes of A. macrocephala ranged from $0.90\;to\;2.35{\mu}m$ with a total length of $16.58{\mu}m$ and the homologous chromosome complement comprised seven metacentrics and five submetacentrics. The total length of A. japonica chromosomes was shorter than that of A. macrocephala, but A. japonica had one subtelocentrics (chromosomes 4) different from A. macrocepha1a. chromosomes. The F1SH technique using 17S and 5S rDNA was applied to metaphase chromosomes. The signals for 17S rDNA were detected on the telomeric regions of chromosomes 4 and 5 in both A japonica and A. macrocephala. The 5S rDNA signal was found in the short arm of chromosome 1.

Discrimination of Species Specific DNA Markers Using RAPD and AFLP Analysis between Atractylodes japonica Koidz. and Atractylodes macrocephala Koidz.

Bang, Kyong-Hwan,Park, Chun-Geon,Jin, Dong-Chun,Kim, Hong-Sig,Park, Hee-Woon,Park, Chung-Heon,Seong, Nak-Sul The Korean Society of Medicinal Crop Science 2003 韓國藥用作物學會誌 Vol.11 No.4

To identify the variation of the RAPD patterns between two Atractylodes species, 52 kinds of random primers were applied to each eight of A japonica and A. macrocephala genomic DNA. Ten primers of 52 primers could be used to discriminate between the species and 18 polymorphisms among 67 scored DNA fragments (18 fragments are specific for A. japonica and A. macrocephala) were generated using these primers, 26.9% of which were polymorphic. RAPD data from the 10 primers was used for cluster analysis. The cluster analysis of RAPD markers showed that the two groups are genetically distinct. On the other hand, to identify the variation of the AFLP patterns and select the species specific AFLP markers, eight combinations of EcoRI/MseI primers were applied to the bulked A. japonica and A. macrocephala genomic DNA. Consequently, three combinations of EcoRI/MseI primers (EcoRI /Mse I ; AAC/CTA, AAC/CAA, AAG/CTA) used in this study revealed 176 reliable AFLP markers, 42.0% of which were polymorphic. 74 polymorphisms out of 176 scored DNA fragments were enough to clearly discriminate between two Atractylodes species.

Genetic Diversity of Rehmannia glutinosa Genotypes Assessed by Molecular Markers

Kyong-Hwan Bang(방경환),Jong-Wook Chung(정종욱),Young-Chang Kim(김영창),Jei-Wan Lee(이제완),Hong-Sig Kim(김홍식),Dong-Hwi Kim(김동휘) 한국생명과학회 2008 생명과학회지 Vol.18 No.4

RAPD 분석을 이용하여 지황 육성 계통과 지역 수집종 들을 구분할 수 있는 분자표지자를 선발하고, 집단 간, 집단 내 유전적 다양성을 평가하기 위하여 본 실험을 수행하였다. 총 20개의 임의 primer를 이용하여 PCR 한 결과, 육성 계통과 수집종 들을 구별할 수 있는 OPA-1 등 10개의 재현성과 다형성이 좋은 프라이머 들을 선발하였다. 특히 OPA-10, OPA-11 및 OPA-19는 고려지황과 지황1호를 다른 계통 및 수집종 들과 구별할 수 있었으며, 이들 프라이머를 이용하여 0.9 kb, 1.2 kb, 1.3 kb 및 1.4 kb 등의 육성계통 특이적인 DNA 밴드들을 확보할 수 있었다. 한편 이들의 결과를 토대로 통계처리에 의한 유전분석 결과, 고려지황, 지황1호 및 일본지황은 집단 내 유사도가 높아 다른 집단들과 구별되었다. 결론적으로, RAPD 분석을 통한 결과는 지황의 유전적 다양성 이해와 특정 계통을 다른 계통 및 수집종 들과 구분할 수 있는 방법으로 이용될 수 있다. Random amplified polymorphic DNA (RAPD) markers were used to identify the genetic diversities among and within varieties and landraces of Rehmannia glutinosa. Polymorphic and reproducible bands were produced by 10 primers out of total 20 primers used in the experiment. In RAPD analysis of the 11 genotypes, 64 fragments out of 73 amplified genomic DNA fragments were polymorphic which represented an average 6.4 polymorphic fragments per primer. Number of amplified fragments with random primers ranged from 2 (OPA-1) to 13 (OPA-11) and varied in size from 200 bp to 1,400 bp. Especially, OPA-10, OPA-11 and OPA-19 primers showed specific bands for varieties of Korea Jiwhang and Jiwhang il ho, which could be useful for discriminating from other varieties and landraces of R. glutinosa. Percentage polymorphism ranged from a minimum of 50% (OPA-1) to a maximum of 100% (OPA-11), with an average of 87.7%. Similarity coefficients were higher in the genotypes of Korea Jiwhang and Jiwhang il ho than in other populations. In cluster analysis, genotypes of Korea Jiwhang, Jiwhang il ho, and Japanese accession were separated from those of other varieties and landraces. Average of genetic diversity within the population (HS) was 0.110, while average of total genetic diversity (HT) was 0.229. Across all RAPD makers the GST value was 0.517, indicating that about 52% of the total genetic variation could be explained by RAPDs differences while the remaining 48% might be attributable to differences among samples. Consequently, RAPD analysis was useful method to discriminate different populations such as domestic varieties and other landraces. The results of the present study will be used to understand the population and evolutionary genetics of R. gllutinosa.

Genetic relationships and molecular authentication of plant origins and the commercial medicinal herbs in peony using RAPD markers

Bang, Kyong-Hwan,Jung, Jin-Ho,Kim, Ok-Tae,Chung, Jong-Wook,Ham, In-Hye,Seong, Nak-Sul,Luo, Rong,Zhang, Gui-Jun,Choi, Ho-Young Kyung Hee Oriental Medicine Research Center 2007 Oriental pharmacy and experimental medicine Vol.7 No.1

Genetic polymorphism and molecular authentication were investigated with the commercial medicinal herb, Peony (Paeonia spp.), using random amplified polymorphic DNA (RAPD) markers. To identify the polymorphism of the RAPD patterns among plant origins, 20 different random primers were applied to the genomic DNA extracted from Paeonia spp. plants such as Paeonia (P.) lactiflora, P. officinale and P. japonica. Ten primers out of 20 primers could be used to discriminate the plant species in the same genus and 72 out of 81 scored DNA fragments (88.9%) generated with these primers were polymorphic. Especially, four primers, such as OPA1, OPA3, OP9, and OPA13, were useful to discriminate the plant origins among the species of Peony. In the results of cluster analysis using RAPD data obtained from the 10 primers, Peony (Paeonia spp.) plants used in this study were grouped into the two distinctive clusters, genetically. Herb medicine, especially P. lactiflora, were easily identified, when species-specific primers were applied to the investigation for discriminating herb medicine currently traded in domestic herb market, Kyungdongmart. Consequently, RAPD analysis was useful method to discriminate plant origins and the commercial medicinal herbs, Paeonia spp..

Comparative Cytogenetic Characteristics and Physical Mapping of the 17S and 5S Ribosomal DNAs between Atractylodes japonica Koidz. and Atractylodes macrocephala Koidz.

Kyong-Hwan Bang,Dal-Hoe Koo,Hong-Sig Kim,Beom-Heon Song,Yong-Gu Cho,Joon-Hyeong Cho,Jae-Wook Bang 한국약용작물학회 2003 한국약용작물학회지 Vol.11 No.4

This study was carried out to compare chromosomal characteristics between Atractylodes japonica and A macrocephala. Cytogenetic analysis was conducted based on karyotype analysis and physical mapping using fluorescence in situ hybridization. As a result of karyotype analysis by feulgen staining, somatic chromosome numbers of A. japonica and A. macrocephala were 2n=24. The length. of the mitotic metaphase chromosomes of A. japonica ranged from 0.70 to 1.60μm with a total length. of 12.11μm and the homologous chromosome complement comprised six metacentrics, five submetacentrics and one subtelocentrics. On the other hand, the length of the mitotic metaphase chromosomes of A. macrocephala ranged from 0.90 to 2.35μm with a total length of 16.58μm and the homologous chromosome complement comprised seven metacentrics and five submetacentrics. The total length of A. japonica chromosomes was shorter than that of A. macrocephala, but A. japonica had one subtelocentrics (chromosomes 4) different from A. macrocepha1a. chromosomes. The F1SH technique using 17S and 5S rDNA was applied to metaphase chromosomes. The signals for 17S rDNA were detected on the telomeric regions of chromosomes 4 and 5 in both A japonica and A. macrocephala. The 5S rDNA signal was found in the short arm of chromosome 1.

우리나라 인삼 육종의 주요 성과와 전망

방경환(Kyong-Hwan Bang),김영창(Young-Chang Kim),이정우(Jung-Woo Lee),조익현(Ick-Hyun Cho),홍지은(Chi-Eun Hong),현동윤(Dong-Yun Hyun),김장욱(Jang-Uk Kim) 한국육종학회 2020 한국육종학회지 Vol.52 No.S

Artificial selection of ginseng has been practiced since Hwangsook (with yellow pericarp and a green stalk, and was developed from a landrace parent) and Cheonggyeong (with red pericarp) were selected as breeding lines in 1926. Systematic research into ginseng breeding, however, started in earnest in the 1960s when the Central Research Institute of Monopoly and Technology (CRIMT) was established, and the Korean Ginseng Experiment Station was organized under the CRIMT. Research into variant characteristics, resource collections, and genetic evaluations began around this time. With the establishment of the Korean Ginseng Institute in the 1970s, studies involving pedigree selection, cataloguing of agricultural traits of genetic resources, generation shortening by tissue culture, and heritability assessments were conducted. In the 1980s, regional adaptation tests were carried out on breeding lines, focusing on ginseng-producing districts. In the 1990s, research was performed on seed multiplication for variety diffusion, effective components and processing quality, and cross breeding. Foreign ginsengs were introduced for interspecies hybridization, and studies were conducted using genetic engineering techniques. Since the 2000s, applications have been made to patent different ginseng cultivars. Currently, 32 cultivars are registered at the Korea Seed & Variety Service. Future goals for ginseng breeding include developing climate change- and disaster-resistant, consumer-oriented, high-performance cultivars. Therefore, it is necessary to develop technologies for distributing new cultivars by collecting and evaluating genetic resources, and cross breeding and performing mass propagation using these resources.