RAPD-PCR(Random Amplified Polymorphic DNAs-Polymerase Chain Reation) 기법에 누에의 유전적 변이 분석을 위한 첫 단계로 다양한 GC함량을 갖는 random primer에 의해서 증폭되는 DNA 단편의 양상 및 증폭도를 비교하였다. RAPD-PCR을 위한 random primer의 증폭도는 GC함량에 의해서 상당히 영향을 받음이 분석되었다. 특히, 50% GC 함량을 갖는 primer는 그 증폭도에 따라서 4가지의 그룹으로 DNA단편이 증폭되었으며 〔bad amplification (75.5%), poor amplification (11.1%), good and excellent amplification(11.1%)〕, primer의 GC 함량이 증가할수록, 휠씬 더 좋은 증폭도를 보여주었다. 그러나, 40% GC 함량을 갖는 primer에 의해서는 어떤 증폭산물도 관찰되지 않았다. PCR을 수행하기 전에 6가지의 제한효소(BamHI, HindIII, Xbal, HaeIII, MspI, Rsal)를 사용하여 누에 genomic DNA를 처리하여 이를 주형 DNA로 하여 RAPD-PCR을 수행한 결과, 유전적 마커의 생산에 대한 효율이 증가함을 알 수 있었다. 이상의 결과를 종합해 볼 때 60%이상의 GC함량을 갖는 random primer와 전처리한 주형 DNA의 사용은 여러 가지 다른 누에 계통의 동정 및 연관군 지도작성에 따른 경비 및 시간을 줄이는데 효율적이라고 사료된다.

Eleven RAPD primers were assessed to analyze genetic diversity of Korean wheat varieties and to develop DNA marker for cultivar identification. The average of the number of polymorphic bands was 5.2 and PIC values showed 0.48, respectively. Ten major clades were presented by phylogenetic analysis. Three cultivars containing Uri, Hanbeak and Jonong were distinct from the others in the phylogenetic dendrogram. Seven cultivar-specific fragments were detected from 11 RAPD fingerprinting among 35 wheat cultivars and they were sequenced. Four Korean wheat cultivars, Eunpa, Jopoom, Yeonbaek and Jeokjoong, were identified newly by four markers, 84, 173, 174 and KWSM011. We convince that these new DNA markers are useful for cultivar fingerprinting and are applied to marker-assisted selection in wheat breeding program.

Background : Cucuma longa L., in the family Zingiberaceae, is distributed in tropical and/or sub-tropical regions mainly in India and China. This species is commonly called tumeric, powder is used as medicinal herbs and/or flavor enhancer. It has been cultivated in southern region mainly Jindo. However, it might be possible to extend cultivation region due to rise in average temperature. In order to select superior lines, agronomic characteristics is commonly used. Because this is not the ultimate solution, the DNA marker approach has benefited the modern plant breeding. Therefore an easy approach by using one kind of primer have been developed from random amplification of polymorphic DNA sequences (RAPD) to discriminate effectively between different cultivars of Cucuma species Methods and Results : DNAs were extracted from the harvested roots of Cucuma sp. using DNeasy plant Mini kit (Qiagen, Hilen, Germany). These plants cultivated from GARES (Hamyang) and used for PCR amplification. The relative concentration of the extracted DNA was estimated Nano Drop ND-1000 (NanoDrop Technologies, Wklmington, De, USA) and final DNA concentration was adjusted to 5.5 ng/㎕. In this study 9 primer pairs were tested on 8 Cucuma sp. These primers showed polymorphism in Cucuma sp. The cluster dendogram showed that the similarity coefficients ranged from 0.68 to 0.87, CUR02 turned out to be CUR11, and CUR16 is similar to CUR17. Conclusion : These finding could be used for further research on cultivar development by using molecular breeding techniques and for conservation of the genetic diversity of Cucuma species. These data on polymorphism difference based on RAPD will be give us invaluable breeding information by selection of superior lines.

Balloon flower (Platycodon grandiflorum A. DC.) is a perennial plant of mainly Campanulaceae family, which have been widely used as a food ingredient and herbal medicine in East Asia. Although demands on related products and yearly cultivation area for balloon flower are increasing, diverse fundamental technologies and molecular breeding studies are not very well supported in Platycodons. In this study, 30 random amplification of polymorphic DNA (RAPD) primers were test in an attempt to explore genetic diversities. In addition, sequences information of the actin gene, a well conserved gene encoding a globular protein that forms microfilaments, was retrieved and analyzed. Two actin homologs were recovered; 3.4 kb fragment is a Pg-actin and 1.4 kb fragment is a Pg-actin homolog with 28.6% similarity. We have confirmed that the Pg-actin gene is configured into 4 exons and 3 introns. A single nucleotide polymorphism (SNP), G↔A, was detected on the intron 3, which served as a target for the CAPS marker development. The marker Pg-Actin-Int3 was applied to 32 balloon flower accessions. Balloon flower DNA sequence information generated in this study is expected to contribute to the analysis and molecular breeding and genetic diversity analysis of balloon flowers.

Angelica gigas, also called Dang Gui or Korean Angelica, is a major medicinal herb used in Asian countries such as Korea, Japan and China. In Korea, we are using the roots of A. gigas., but, they are using Angelica sinensis in China and using Angelica acutiloba. in Japan to obtain many active constituents such as dercursin, decursinol angelate, nodakenetin, nodakenin, umbelliferone, β-sisterol, or α-pinene. The plants of the Angelica family are used to improve gynecological health. The biggest problem in the cultivation of A. gigas is bolting. If the bolting occurs, A. gigas can not be used as a medicinal component because the roots are lignified. In this study, 11 A. gigas genetic resources in Korea; 1. Hwangje variety, 2. Sungwoo Jongmyo company, 3. Bonghwa No. 1, 4. Bonghwa No. 2, 5. Bonghwa No. 3, 6. Bonghwa No. 4, 7. Jechun local variety, 8. Jirisan local variety, 9. Manchu variety in Eumseong, 10. Manchu variety in Bonghwa, 11. Jinbu local variety, were collected and performed phylogenetic analysis using RAPD molecular markers.

Adenophora triphylla var. japonica HARA is a herbaceous plant belongs to Campanulaceae. Adenophora root is mainly used for medicinal purpose. It is effective for lung cleaning, sputum remove, viscera strengthening, cough stopping and cancer treatments. Adenophora has about 70 species in the world and 17 of the species are distributed in Korea. Genetic resources of A. triphylla var. japonica HARA are valuable as the habitat is concentrated in East Asia. The intraspecies variation is very high according to the environmental conditions. A new A. triphylla var. japonica HARA variety, ‘Harang’, was developed through polyploid breeding in 2011. But, low domestic production and passive studies caused our country to rely on imports for almost all amount of the A. triphylla var. japonica HARA demands. In this experiment, genetic diversity between the collections were analyzed using 32 RAPD primers. Through this study, limit of morphologic classification could be solved and genetic diversity of this plant could be assured.

Codonopsis lanceolata is used as a natural medicine or vegetables. It originates in East Asia such as Korea, Japan and China. Similar to Panax ginseng, C. lanceolata contains saponins as effective components. C. lanceolata is cultivated in many regions of South Korea. But, no variety was developed yet and the origin discrimination in the distribution market of C. lanceolata became a problem. In this study, we collected 20 C. lanceolata regional groups; Hoengseong, Wonju, Samcheok, Chuncheon, Pyeongchang, Hongcheon, Yongin, Yangpyeong, Danyang, Chungju, Bonghwa, Ulleung, Yeongju, Sancheong, Muju, Gwangyang, Sinan, Hwasun, Jeju-si and Seogwipo-si, and tested the genetic relationship using RAPD molecular markers. The genomic DNA was extracted using CTAB and the RAPD analysis was performed using 32 primers of Operon Technologies. NTsys-PC program was used for the phylogenetic analysis of the data.

Platycodon grandiflorum is a perennial herbal plant belongs to Campanulaceae family. It has very important genetic value as a major plant in Asterids order. The major ingredients are platycosides, terpenoid saponins. In Korean industrial plants market, it was produced 5,633 tons in 2013, and the total amount of production was less than only five species, omija, ginger, raspberry, yam and deodeok. P. grandiflorum is called ‘Gilgyung’ and is used as a fresh vegetable and an ornamental plant. Nowadays, various components of P. grandiflorum were already published. But, genetic research is in the starting stage. In this study, 11 cultivars; 1. MariesⅡ, 2. Hakone double white, 3. Hakone double blue, 4. Fuji white, 5. Fuji pink, 6. Fuji blue, 7. Astra white, 8. Astra pink, 9. Astra blue, 10. Astrasemi double blue, 11. Jangback, were analyzed using 60 Operon Universal RAPD primers. The results were phylogenetically analyzed and related to the morphological characteristics of the cultivars.

김정문 알로에 제주농·공장에서는 알로에 베라와 알로에 아보레센스를 중심으로 450여종의 알로에가 보존되어 져 오고 있으며, 알로에 종 일부는 전시용으로도 활용되고 있다. 전세계 500여종의 알로에가 알려져 있는 가운데 제주농·공장의 알로에 보존 현황은 원산지가 아님에도 불구하고 많은 종수를 보유하고 있으며 보존 또한 잘 이루 어지고 있다. 또한 변종 및 아종과 더불어 교잡종 등의 분류가 제대로 이루어진다면 분류군은 증가하리라 예상된 다. 따라서 제주농·공장에 보존되고 있는 다양한 알로에 종을 김정문 알로에의 경제적 자원으로서 활용하기 위해 서 이들의 유연관계를 분자생물학 기술을 활용하여 분석하고 품종 구분에 대한 기술을 확보하는 것이 필요하다. 이에 따라 김정문 알로에 제주농·공장에 보존되어 있는 품종 중 경제성이 있을 것으로 판단되는 35종의 품종을 대 상으로 하여 Operon 사의 OPF 프라이머 5종, OPD 프라이머 15종을 사용하여 RAPD 분석을 수행하였다. 이를 통해 품종별 유연관계를 분석하였고, 10개의 그룹으로 알로에가 분류될 수 있음을 확인하였다. 이와 같은 연구를 통해 품종판별용 RAPD 마커 제작도 가능할 것으로 기대된다.

Chrysanthemum white rust, caused by Puccinia horiana, is one of the most destructive fungal diseases in chrysanthemum cultivation worldwide. For increasing efficiency of resistant breeding, molecular markers linked to chrysanthemum white rust resistance gene were developed in pseudo F1 cross population between ‘Puma White’ as susceptible and ‘Dancer’ as resistant using bulked segregant analysis (BSA). Of 280 RAPD primers (Operon 10 mer), 18 primers found to be polymorphic. After screening of these primers in 20 individual lines, only OPI-13520 was selected as closely linked marker to white rust disease resistance. Based on correspondence between phenotypic resistant level and marker in 187 segregation population, the genetic distance between white rust resistance gene and OPI-13520 marker assumed to be 3.8 cM. For OPI-13520 marker conversion into sequence characterized amplified region (SCAR) marker, the amplified fragment of OPI-13520 was purified, cloned and sequenced. Based on the DNA sequence of OPI-13520, SCAR maker was generated and verified in 20 individual lines used in BSA-RAPD.The results showed SCAR marker could be used to identify white rust resistance in chrysanthemum.

The fruits of Schisandra chinensis have been used as an edible ingredient and traditional medicine in Korea. Due to morphological similarities of dried mature fruits, the correct identification of S. chinensis from other closely related Schisandrae species is very difficult. Therefore, molecular biological tools based on genetic analysis are required to identify authentic Schisandrae Fructus. Random amplifed polymorphic DNA (RAPD) and Sequence Characterized Amplified Region (SCAR) were used to develop an easy, reliable and reproducible method for the authentication of these four species. In this paper, we developed several RAPD-derived species specific SCAR markers and established a multiplex-PCR condition suitable to discriminate each species. These genetic markers will be useful to distinguish and authenticate Schisandrae Fructus and four medicinal plants, S. chinensis, S. sphenanthera, S. repanda and K. japonica, in species level.

This study was conducted to investigate the environmental characteristics of natural habitats, and the genetic variations in samples from 19 different quadrates of Asplenium scolopendrium 10 habitats. The natural habitats of A. scolopendrium were located at an altitudes 110-973 m with inclinations of 7-30˚. All the quadrates were located on north facing slopes. The average field capacity of the soil is 31.4%, with organic matter at 16.7%, and the pH is 5.84. The soil texture was confirmed as sandy loam in 8 habitats and silt loam in 2 habitats. A total of 214 vascular plants were identified from 10 habitats. The importance value of the herbaceous layer (H) was the highest in A. scolopendrium, at 10.4%, followed by Arachniodes standishii (7.3%), Dryopteris crassirhizoma (6.5%), and Polystichum tripteron (5.3%), which implies that the natural habitats of A. scolopendrium are affinity with ferns. The species diversity of A. scolopendrium was estimated as on average 1.09, while the dominance and evenness were 0.12 and 0.87, respectively. The result of the RAPD analysis, among 59 bands amplified with a primer, 25 (42.4%) showed polymorphism. Twenty-one individuals of 10 habitats could be classified into four groups with similarity coefficient values ranging from 0.74 to 1.0. Mt. Mullae and Geumdaebong populations shows basal branching within the 21 individuals. Ulleung-do island and Jeju-do island population forms an independent clade, respectively. Mt. Moak and Byeonsan-bando clade formed a sister to the Ulleung-do island and Jeju-do island clade.

포도 대목 품종 육성 시, 주요 교배친으로 사용되는 대목 품종 29점 및 국내 자생 머루를 포함한 포도 속(Vitis) 야생 유전자원 13점 등 총 42점의 유전적 다양성을 분석하기 위하여 RAPD와 SSR 분석을 수행하였다. Random decamer 30 종을 분석하여 329개의 다형성 밴드(60.3%)를 얻었으며 primer 당 평균 다형성 밴드 수는 11.0개였다. SSR 마커 20종을 이용하여 분석한 결과 263개의 대립인자가 확인되었고, 마커

Mulberries have importance in the sericulture industry as food for Bombyx mori, silkworm reared for its silk. Korean Morus alba have many cultivars and, for the protection of these cultivars and for utilization in plant-breeding programs, genetic information and the diversity among cultivars are essential. This study with 14 mulberry genotypes was undertaken using RAPD and ISSR fingerprinting to discover the genetic divergences between cultivars. Polymorphism rate among the cultivars produced by RAPD primer was found to be 64.48% and 66.29% relative to ISSR primer. The genetic relationships among the cultivars were identified using a dendrogram constructed with the UPGMA clustering method. Nei's method was used to calculate the genetic dissimilarity coefficients between each pair of genotypes, and the highest dissimilarity coefficient of 0.246 was exhibited between Suwon and Hwanggum cultivars. To determine the efficiency of each primer, a polymorphic index was calculated, and the robustness of the dendrogram was checked using cophenetic correlation coefficient. The results of this study can be utilized for the improvement of mulberry varieties in plant-breeding programs.