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연구논문 : 자연과학; RAPD 마커를 이용한 인삼 품종 및 육성계통의 유전적 다양성 분석
방경환 ( Kyong Hwan Bang ),정종욱 ( Jong Wook Chung ),김영창 ( Young Chang Kim ),조익현 ( Ick Hyun Jo ),김장욱 ( Jang Uk Kim ),신미란 ( Mi Ran Shin ),현동윤 ( Dong Yun Hyun ),김동휘 ( Dong Hwi Kim ),차선우 ( Seon Woo Cha ),김기홍 한국국제농업개발학회 2013 韓國國際農業開發學會誌 Vol.25 No.2
RAPD 마커를 이용하여 인삼 품종 및 육성계통의 유전적 다양성 및 유연관계를 분석한 결과는 다음과 같다. Fig.5. Dendrogram of genetic similarity among Korean ginseng cultivars and breeding lines constructed on the data obtained with 25 RAPD primer 1. 총 130개의 primer 중 polymorphism을 나타내는 70개의 primer를 선발하였고, 그 중 재현성이 있으면서 polymorphism 이 높은 25개의 primer를 선발하였다. 증폭된 DNA 단편의 수는 189개이고, PCR 산물은 100 ~ 2,800 bp 범위로 증폭되었다. 2. 각 primer에 의해 증폭된 DNA 단편의 수는 3개 ~ 17개로 다양하였으며, primer 한 개당 평균 7.6개의 DNA 단편이 증폭되었다. OPD19 primer를 이용한 유전분석 결과, 총 5개의 유전양상이 나타났는데, 약 500 ~ 1,300 bp의 증폭산물에서 품종 및 계통 간 유전적 다형성을 나타냈다. 3. 선발된 primer별 대립인자는 최소 1.33에서 최대 2.00의 범위였고, 평균 1.709이었다. primer별 유전적 다양성은 OPD15가 가장 높았고, OPF2가 가장 낮은 값을 나타내었다. 본 연구에서 분석에 이용된 25개의 RAPD primer 중에서 D15, D19, B5, A19등은 인삼 품종과 계통에서 비교적 높은 수의 대립단편과 높은 유전적 다양성 값을 나타내는 primer 였다. 4. 유사도 계수 0.98을 기준으로 24개의 품종 및 계통을 대상으로 군집분석을 수행한 결과, 미국에서 수집 육성된 G04116과 국내 품종인 천풍, 연풍 그리고 국내 육성 계통인 G04009, G04026, G04069, G04084는 그룹을 형성하지 않았고, 17개의 품종 및 계통은 2그룹으로 분류되었다. I 그룹에는 고풍, 금풍과 12계통(85%), II 그룹에 3계통(15%)이 포함되었다. In this study, Random Amplified Polymorphic DNA (RAPD) analyses were used to clarify the genetic polymorphisms among Korean ginseng cultivars and breeding lines and to classify them into distinct genetic groups. Among 130 primers in RAPD analysis, 25 primers were selected as the appropriate identification of genetic characteristics in Korean ginseng cultivars and breeding lines. Twenty-five RAPD loci generated a total of 189 DNA bands. Amplified PCR products were showed the highly reproducible banding patterns at 100 ~ 2,800 bp. The number of amplified bands for each RAPD primers ranged from 3 to 17 with a mean of 7.6 bands. In the case of OPD19 primer, 5 banding patterns were displayed in Korean cultivars and breeding lines. Banding patterns were approximately 500 to 1,300 bp amplicons. The number of alleles for each RAPD locus ranged from 1.33 to 2.00 with a mean of 1.71 alleles. OPD15 and OPF02 primers demonstrated the highest and the lowest genetic polymorphisms, respectively. Cluster analysis based on genetic similarity estimated by RAPD markers classified Korean cultivars and breeding lines into 2 groups. Group I included Gopoong and 11 breeding lines (50%); group II included 3 breeding lines (1.3%). Consequently, the results of ginseng-specific RAPD markers may prove useful for the evaluation of genetic diversity and differentiation of Korean ginseng cultivars and breeding lines.
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.
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.
인삼 약 배양을 통한 배 발생과 식물 재분화 및 유세포 분석기를 이용한 배수성 검정
이정우,방경환,김동휘,김장욱,김영창,조익현,Jung-Woo Lee,Kyong-Hwan Bang,Dong-Hwi Kim,Jang-Uk Kim,Young-Chang Kim,Ick-Hyun Jo 한국식물생명공학회 2023 JOURNAL OF PLANT BIOTECHNOLOGY Vol.50 No.1
이번 연구에서 인삼의 약 배양을 통한 배 형성과 식물체의 재분화가 확인되었다. 인삼의 약 배양에서 캘러스 유도에 적합한 기본 배지는 MS 배지였고 sucrose 농도는 3%였으며, 배 형성에 적합한 호르몬 농도는 2,4-D 1.0 mg/L 또는 3.0 mg/L이었다. 저온 전처리에 의한 인삼 약 배양의 캘러스 유도와 배 형성의 효율성 차이는 확인할 수 없었다. 인삼의 품종별 약 배양의 효율성을 검정한 결과, 청선 품종의 배 형성률이 가장 우수하였으며 선운 품종은 배 형성이 관찰되지 않았다. 약 배양으로 유도된 캘러스의 배수성을 검정한 결과, 반수체 캘러스가 확인되었지만 이 캘러스에서 배는 형성되지 않았다. 약 배양 유래의 배를 발아배지에 치상하였을 때 정상적으로 식물체가 재분화 하였으며, 이 식물체의 배수성을 검정한 결과, 모두 2배체로 확인되었다. 추후 DNA 마커를 이용한 약 배양 유래 식물체의 동형접합성의 확인이 필요하다고 판단된다. 인삼 약 배양을 통하여 동형접합성 식물체를 생산할 수 있다면 인삼 육종 효율성을 증진시키는데 크게 기여할 수 있을 것이다. Korean ginseng (Panax ginseng Meyer) is an economically important plant because of it is rich in saponins. It is mainly cultivated in Asia, including Korea and China. Since ginseng requires a long breeding period due to juvenility, homozygote production techniques, such as anther culture, must be urgently established. In the present study, callus induction and embryogenesis through anther culture were observed in P. ginseng. Murashige and Skoog medium was used as the basal medium suitable for callus induction. When the medium was supplemented with 3% sucrose, the callus induction rate was high and the callus size was large. Cold pretreatment did not significantly affect callus induction and embryogenesis. Embryogenesis was the most efficient when the embryo-formation medium was supplemented with 1.0 or 3.0 mg/L 2,4-dichlorophenoxyacetic acid. Cultivar significantly affected anther culture efficiency. Specifically, 'Cheongseon' showed the highest embryo-formation efficiency, whereas no embryogenesis occurred in 'Sunun'. Ploidy assessment revealed the haploid status of the induced calli. Embryos derived from anther culture formed shoots upon transfer to germination medium, although no difference in ploidy was noted between the induced callus and control. Overall, the anther culture conditions established in the present study may contribute to the production of homozygous P. ginseng plants in the future.
Multicolor FISH와 Feulgen 염색법을 이용한 Angelica속 식물의 세포유전학적 분석
구달회,김수영,방경환,성낙술,방재욱,Koo, Dal-Hoe,Kim, Soo-Young,Bang, Kyong-Hwan,Seong, Nak-Sul,Bang, Jae-Wook 한국식물생명공학회 2003 식물생명공학회지 Vol.30 No.2
Karyotype analysis and chromosomal localization of 5S and 45S rDNAs using multi-color fluorescence in situ hybridization (McFISH) technique were carried out in two Angelica species. The numbers of diploid chromosomes were the same in two same in two species as 2n=22, however the lengths of chromosomes were varied from 4.25 to 6.50 ${\mu}{\textrm}{m}$ in A gigas and 4.95 to 8.50 ${\mu}{\textrm}{m}$ in A acutiloba. The chromosomes of A. gigas were composed of five metacentric and six submetacentric pairs, while those of A. acutiloba were six metacentic, one submetacentric and four subtelocentric paris. In FISH experiments, the numbers and size of 45S rDNA signals were varied between two species, however dach signal of the 5S rDNA was observed in two species.
인삼 종자의 개갑률 향상을 위한 적정 수확시기 및 GA<sub>3</sub> 처리 효과
김영창,김영배,박홍우,방경환,김장욱,조익현,김기홍,송범헌,김동휘,Kim, Young Chang,Kim, Young Bae,Park, Hong Woo,Bang, Kyong Hwan,Kim, Jang Uk,Jo, Ick Hyun,Kim, Kee Hong,Song, Beom Heon,Kim, Dong Hwi 한국약용작물학회 2014 韓國藥用作物學會誌 Vol.22 No.6
This study was performed to identify optimal harvesting time of ginseng seeds and to examine the effect of $GA_3$ treatment for improvement of seed stratification rate. Ginseng seeds harvested from Land race, Chunpoong and Yunpoong cultivar in July 20 were tested for stratification rate. It was shown that stratification rates of land race, Yunpoong and Chunpoong cultivar were 94.1%, 93.1%, and 82.6%, respectively. Seeds of Chunpoong cultivar harvested 10-15 days later showed a comparable stratification rate to that of Land race, indicating that late harvest of Chunpoong seeds is beneficial for the increase of stratification rate. The higher stratification rate was found in mature seeds (92.3%) than immature seeds (37.8%), both of which were harvested in July 20. Stratification rate of mature seeds harvested in July 15 was 87.5%, demonstrating optimal harvesting time of ginseng seeds with higher stratification rate is after mid-July. An exponential growth of endosperms of ginseng seeds was observed from early June to mid-June and then slow growth was observed. There was no obvious growth of embryos from fertilization to mid-August. After the this time, embryos quickly grew until late October. Thus, appropriate stratification control is essential during the period (from early September to late October) in order to optimize embryo growth and development. While no increase of stratification rate was observed in seeds treated with 50 ppm of $GA_3$, significant increases were observed in seeds treated with 100 ppm of $GA_3$. At this concentration of $GA_3$, the stratification rate of Land race, Chunpoong and Yunpoong cultivar was 95.0%, 95.3%, and 96.5%, respectively.