EST-SSR 마커를 이용한 인삼 품종과 육성계통의 유전적 다형성 및 유연관계 분석

방경환,서아연,정종욱,김영창,조익현,김장욱,김동휘,차선우,조용구,김홍식 韓國藥用作物學會 2012 한국약용작물학회지 Vol.20 No.4

In this study, Expressed Sequence Tag-Simple Sequence Repeat (EST-SSR) analyses were used to clarify the genetic polymorphisms among Korean ginseng cultivars and breeding lines and to classify them into distinct genetic groups. Polymorphic and reproducible bands were produced by 14 primers out of total 30 primers used in this study. Fourteen EST-SSR loci generated a total of 123 bands. Amplified PCR products showed the highly reproducible banding patterns at 110~920 bp. The number of amplified bands for each EST-SSR primers ranged from 2 to 19 with a mean of 8.8 bands. P26 and P35 primers showed 13 and 12 banding patterns, respectively. The number of alleles for each EST-SSR locus ranged from 1.67 to 2.00 with a mean of 1.878 alleles. P34 and P60 primers showed the highest and the lowest genetic polymorphism, respectively. Cluster analysis based on genetic similarity estimated by EST-SSR markers classified Korean cultivars and breeding lines into 4 groups. Group included Gopoong and Chunpoong and 9 breeding lines (55%), group included 2 breeding lines (10%), group included 3 breeding lines (15%), group included Gumpoong and 3 breeding lines (20%). Consequently, the EST-SSR marker developed in this study may prove useful for the evaluation of genetic diversity and differentiation of Korean ginseng cultivars and breeding lines.

해부형태적 특징과 SCAR Marker를 이용한 백출의 기원식물 판별

방경환,성정숙,박충헌,김동순,박춘근,유홍섭,박희운,성낙술 韓國藥用作物學會 2004 한국약용작물학회지 Vol.12 No.1

국내 한약재 시장에서 유통되고 있는 백출 건재약재의 기원을 판별하고자 해부형태적 특징과 종 특이적인 SCAR 마커 (SAjR2, SAmR1)를 적용한 실험 결과는 다음과 같다. 해부형태적 특징으로 A. japonica에서는 주피에 코르크층이 매우 발달하여 있었고, 코르크층 내부 또는 아래에 석세포대가 연속적으로 발달하고 있었다. 유실은 A. japonica에서 뿌리횡단면에 전체적으로 다수 분포하여 잘 발달하고 있었으며 피충부분과 잘 발달된 방사조직 내에서도 목섬유다발이 다수 관찰됐다. 또한 유세포 내의 침상 결정물도 A. macrocephala보다 많이 함유하고 있었다. 이러한 특징들을 종합한 결과, 국내 유통 국산 및 수입 백출은 모두 A. japonica의 해부형태적 특징을 따르고 있어 A. japonica로 동정되었다. 한편 RAPD clone으로부터 개발된 A. japonica 특이적인 SAjR2를 이용하여 PCR 한 결과 모든 샘플에서 600bp 크기의 한국백출 특이적인 밴드가 형성되었으며, A. macrocephala 특이적인 SAmR1을 이 용한 결과에서는 작물시험장에서 재배중인 A. macrocephala에서만 1200 bp크기의 특이적인 밴드가 형성되어, 시중에서 유통되고 있는 한국백출과 중국백출은 모두 A. japonica로 동정되었다. 결론적으로 해부형태적 특성과 SCAR 마커는 A. japonica와 A. macrocephala 기원식물의 동정과 유통 건재 약재를 판별하기에 유용한 방법이었다. Finding a means to discriminate the commercial herb medicines when they were dried and sliced is a very important and imminent project in Korea. To differentiate plant origins and the commercial herb medicines of Atractylodes japonica and A. macrocephala, two discriminative methods using anatomical characteristics and SCAR marker were applied. It was possible to discriminate plant origins and the commercial herb medicines between A. japonica and A. macrocephala by anatomical characteristics: development of periderm, layer of stone cell, distribution of laticiferous vessels, development of xylem fiber in xylem ray, contained quantity of clustered crystals and others. While, two SCAR markers were developed from RAPD clones: SAjR2 (600 bp) from AjR2 and SAmR1 (1,200 bp) from AmR1. These two markers were enough for discrimination plant origins and the commercial herb medicines between A. japonica and A. macrocephala. The result of application of anatomical characteristics and SCAR markers to investigate current status in domestic herb market, Daegu and Kumsan herb market, it was identified to be current herb medicines of A japonica.

SSR 마커를 이용한 국내산 인삼 품종 및 국외 수집종의 유전적 다양성 분석

방경환,조익현,정종욱,김영창,이제완,서아연,박종현,김옥태,현동윤,김동휘,차선우 韓國藥用作物學會 2011 한국약용작물학회지 Vol.19 No.5

In this study, simple sequence repeat (SSR) analyses were utilized for evaluation of genetic diversity and discrimination of 17 accessions. Five cultivars, which were developed from Korea, and 12 foreign accessions, which were collected from China, Japan, Russia and USA, were evaluated by nine markers out of 22 SSR markers. A total of 39 alleles were detected, ranging from 2 to 8, with an average of 4.3 alleles per locus. The expected heterozygosity and PIC values were 0.627 and 0.553, with a range from 0.21 (GB-PG-078) to 0.76 (GB-PG-142) and from 0.19 (GB-PG-078) to 0.70 (GB-PG-142), respectively. Four makers out of nine SSR markers, GB-PG-026, GB-PG-043, GB-PG-142 and GB-PG-177, were selected as key factors for discrimination of Korean ginseng cultivars and foreign accessions. All of Korean ginseng cultivars and foreign accessions were individually by the combination of four SSR markers. Consequently, the SSR markers developed in this study may prove useful for the evaluation of genetic diversity and discrimination of Korean ginseng cultivars and foreign accessions.

SSR 마커를 이용한 고려인삼 품종 판별기술 개발

방경환,정종욱,김영창,이제완,조익현,서아연,김옥태,현동윤,김동휘,차선우 韓國藥用作物學會 2011 한국약용작물학회지 Vol.19 No.3

The principal objective of this study was to develop a discrimination method using SSR markers in Korean ginseng cultivars. Five cultivars--Chunpoong, Yunpoong, Gopoong, Sunpoong, and Kumpoong--were evaluated by nine markers out of 22 SSR markers. A total of 23 alleles were detected, ranging from 1 to 4, with an average of 2.6 alleles per locus, and an averages of gene diversity (GD) of 0.480. Nine markers were tested in order to distinguish among five Korean ginseng cultivars. Two markers out of nine SSR markers, GB-PG-065 and GB-PG-142, were selected as key markers for discrimination among Korean ginseng cultivars. Two genotypes were detected in GB-PG-065. Chunpoong and Kumpoong shared the same allele type, and Yunpoong, Gopoong, and Sunpoong shared another identical allele type. In the case of GB-PG-142, a specific allele type differentiated from those of other four cultivars was observed only in Sunpoong cultivar. Consequently, the SSR markers developed in this study may prove useful for the identification of Korean ginseng cultivars and the development of ginseng seed management systems, as well as tests to guarantee the purity of ginseng seeds.

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

방경환,구달회,허만규,성낙술,방재욱 한국유전학회 2004 Genes & Genomics Vol.26 No.1

PNA 바이오칩을 이용한 고려인삼 품종 판별

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.

Random amplified Polymorphic DNA 마커를 이용한 고려인삼 품종과 육성계통의 유전적 다양성 분석

방경환,김영창,조익현,신미란,김장욱,현동윤,김동휘,차선우 한국약용작물학회 2013 한국약용작물학술대회 발표집 Vol.2013 No.05

감마선 처리가 인삼 부정배에 미치는 영향

방경환,이정우,김장욱,조익현 한국약용작물학회 2018 한국약용작물학술대회 발표집 Vol.2018 No.10

Background : Mutagenesis is one of the most powerful method for genetic variation, however there is little research history in P. ginseng. The seeds and vegetative tissues are materials commonly used for mutagenesis, but there is a problem that chimeras known as sectoral differences can occur and unintentional poor selection can occur owing to the influence of the growth environment. The combination of in vitro culture and mutagenesis can eliminate the effects of the environmental factors. Methods and Results : Gamma irradiation was carried out using a 60Co irradiator (3000Ci, Nordion Inc., Ottawa, Canada) of the Korea Atomic Energy Research Institute, Jeongeup, Korea. The explants containing the somatic embryos were treated with 20, 40, 60, 80, and 100 Gy. Gamma-rays affected the somatic embryos produced in vitro depending on the dose. The survival rate of somatic embryo was significantly influenced by gamma irradiation. Also, the development of the embryo was changed by gamma irradiation depending on each dose. When somatic embryos were transferred to the medium supplemented with GA3, they started to germinate but were affected significantly by gamma irradiation dose. When germinated shoots were transferred to 1/2 SH medium to induce rooting, as the concentration of gamma ray treatment increased, the rooting gradually decreased. Conclusion : Considering the overall survival rate, germination rate, embryo development, and rooting rate, the optimal dose of gamma rays to be used with somatic embryo is estimated to be between 60 Gy and 80 Gy.

차세대 염기서열 분석을 이용한 고려인삼과 미국삼의 전사체 특성 비교

방경환,조익현,이승호,김영창,김장욱,신미란,김영배,김동휘,김기홍,김선태 한국약용작물학회 2014 한국약용작물학술대회 발표집 Vol.2014 No.05

홍화의 작물학적 특성에 의한 품종군 분류

방경환,김영국,박희운,성낙술,조준형,박상일,김홍식 한국약용작물학회 2001 한국약용작물학회지 Vol.9 No.4

홍화 수집종들의 작물학적 특성을 통하여 유전적 다양성 및 유연관계를 밝히고, 품종군을 분류하여 품종육성의 기초자료로 이용코자 시험한 결과를 요약하면 다음과 같다. 개화일수는 74~88일 분포로 평균 83일이었고, 80일 이하의 조숙성을 가진 수집종은 21개 이었으며, 1차분지수와 2차분지수는 각각 3.8~14.8개와 0~26.9개의 범위로 분포하였고, 다분지형의 특성을 가진 품종은 IT201434, IT202723 등 이었으며, 생육특성 중 2차 분지수의 변이가 켰다. 수량구성 요소로서 두상화수는 5.0~40.7개, 두상화당 종실수는 25~68개, 100립중 및 주당 종실중은 각각 2.9~5.4g, 3~47.5g의 범위로 분포하였고, 종실중 〉 두상화수 〉 두상화당 종실수 순으로 변이가 큰 경향이었다. 수집종들은 작물학적 특성에 따라 11개군으로 분류되었으며, I군은 25%, II군은 33%, III군은 14%, IV군은 8%, VI군은 3%, IX군은 2%, X군은 1%, ?군은 1%가 속하였으며, III군은 모두 국내재래종 이었다. 분류된 군중 X군은 출아소요일수가 짧고, 개화소요일수가 길었으나 수량 구성 요소인 두상화수, 주당 종실중 및 두상화당 종실수가 가장 우세한 군으로 분류되었다. 수량구성요소인 두상화수와 주당 종실중은 경직경, 제1차분지수, 제2차분지수, 엽수 및 엽장과 높은 정의 상관을 보였다. This study was conducted to provide the basic information on safflower collections and to identify the variations which could be utilized in safflower breeding programs. The agronomic characteristics was used to clarify the genetic relationships among safflower collections and to classify them into distint genetic groups. There were 21 early maturing collections with less than 80 days in number of days from planting to flowering. The number of primary and secondary branches ranged 3.8~14.8 and 0~26.9, respectively, and two collections, IT201434 and IT202723, were found to be high branch types. The 101 safflower collections were classified into 11 groups based on the complete linkage cluster analysis using agronomic characteristics. The I, II, III, IV, IX, X and ? groups included the 25%, 33%, 14%, 8%, 2%, 1% and 1% of the collections, respectively. All the collections in the group III were Korean landraces. The collections in group X could be characterized as early emergence, late flowering and high yield components such as the number of capitula per plant, number of seeds per capitula and seed weight per plant. The number of capitula per plant and seed weight per plant, i.e., the two main yield components, had the highly significant positive correlations with stem diameter, number of the primary branches, number of the secondary branches, number of leaves and leaf length.