본 연구는 한국, 중국, 엘살바도르 강낭콩 유전자원을 대상으로 농업특성을 조사하고, 분자마커를 이용하여 유전적 다양성을 분석하였으며 결과는 다음과 같다. 첫째, 개화일수 전체 평균은 61일, 범위는 41일에서 83일, 생육일수는 평균 104일, 범위는 86일에서 143일 사이였다. 엘살바도르 자원의 평균 생육일수는 97일로 한국, 중국의 104일과 103일보다 6∼7일 빨랐다. 둘째, 협장은 전체 평균 10.9 cm, 범위는 4.2∼19.5cm, 중국 자원의 평균 협장은 12.7 cm로 가장 길고 범위도 7.0∼19.5 cm로 넓었다. 협폭의 전체 평균은 11.7 mm, 범위는 5.6∼27.5 mm, 중국 자원의 평균 협폭은 12.8 mm, 범위는 6.7∼27.5 mm로 3개국 중 가장 넓었다. 셋째, 100립중의 전체 평균은 40.9 g, 범위는 18.1∼69.3 g, 한국과 중국 자원의 평균 100립중은 각각 44.3 g, 42.7 g으로 유사하였으나 엘살바도르 자원의 평균 100립중은 26.3 g으로 매우 작았으며 분포범위도 18.5∼32.1 g으로 좁았다. 넷째, 신육형은 유한형이며 직립형 자원이 35.7%, 무한형이며 덩굴성인 자원이 46.4%였으며, 신육형이 중간형이고 덩굴성 자원이 16.1%로 나타났다. 한국 자원은 유한형과 무한형의 비율이 유사하였으며, 중국 자원의 신육형은 무한형이며, 덩굴성 자원이 60.5%, 엘살바도르는 중간형이며, 덩굴성 자원이 90.1%로 대부분을 차지하였다. 다섯째, 종피색은 전체적으로 적색자원이 평균 44.5%로 가장 많았으며, 특히 엘살바도르는 적색자원이 83.3%로 대부분이었고, 종자모양은 신장형이 평균 30.4%로 가장 많았으나, 한국자원은 계란형(36.6%), 중국자원은 신장형(55.3%), 엘살바도르는 입방형(79.2%)이 많아 국가별 차이를 보였다. 여섯째, 한국 자원 292점, 중국 자원 72점, 엘살바도르 자원 71점 등 435점을 대상으로 8개의 SSR 마커를 이용한 유전적 다양성 분석을 수행하였으며, 그 결과 총 92개의 allele가 확인되었고, gene diversity 와 PIC로 볼 때 중국 자원이 각각 0.73, 0.69로 가장 높았으며, 엘살바도르 자원은 0.48과 0.45로 가장 낮은 것으로 나타났다.

In this study, genetic diversity of wild Codonopsis lanceolata collected in Korea were analysed using SSR makers. Wild C. lanceolata roots were collected in Jeollanam-do Jangheung-gun Choentae Mountain as in roots. The wild C. lanceolata plants were cultivated in Chungbuk National University greenhouse and the leaves were sampled from 36 plants. The genomic DNA of C. lanceolata was extracted using CTAB. PCR was performed using a program of 35 cycles at 94℃ for 30 sec, 60℃ for 30 sec, and 72℃ for 30 sec with an pre-denaturation of 94℃ for 5 min and a final extension of 72℃ for 30 min. The PCR reaction mixture contains 5 pmole of primers and 20 ng of DNA template in a 20 μL reaction volume. The genotype of the analyzed samples were very different. Therefore, the wild C. lanceolata collected in Korea look genetically diverse.

Codonopsis lanceolata is a perennial climber. The roots are used as medicinal materials or vegetables. Recently, demand for C. lanceolata is increasing as a healthy food. C. lanceolata is distributed in India and East Asia such as China, Japan as well as Korea. In South Korea, this plant is widely cultivated in Gangwon-do province. No C. lanceolata varieties were developed in Korea. The objective of this study is to analyze genetic diversity of C. lanceolata cultivated in Korea using SSR makers. C. lanceolata roots were collected in each region were cultivated in Chungbuk National University greenhouse. Samples were obtained from fresh leaves of 5 plants from each collection region. The genomic DNA was extracted using CTAB. Genetic diversity was analysed using 4 sets of C. lanceolata SSR makers. PCR was performed in total 20 μL reaction volume containing 20 ng of DNA template, 5 pmole of primers. The genotypes of the analyzed samples were very similar. That means that the genetic diversity of C. lanceolata cultivated in Korea is very low.

Blueberry (Vaccinium spp.) is a member of the Ericaceae and eleven varieties have been registered at the Korea Seed & Variety Service for Plant Variety Protection (PVP). This study was to develop simple sequence repeat (SSR) markers next generation sequencing (NGS) analysis and to analysis genetic relationship of blueberry 31 varieties. Highbush blueberry ‘Camellia’ and rabbiteye blueberry ‘Alapaha’ varieties were used as sequencing materials. Out of total 987 SSR primers detected between ‘Camellia’ and ‘Alapaha’, 148 SSR primers were initially applied to select SSR markers for identification of blueberry varieties. Fourteen SSR markers showed polymorphism between 8 varieties. Seven SSR markers showed reproducibility and clear peak among 14 SSR markers. Genetic relationships of 31 blueberry varieties were analyzed and identified using 7 SSR markers. A total of 30 polymorphic SSR alleles were obtained and two to seven alleles were detected for each locus with an average of 4.3 alleles per locus. Average polymorphism information content was 0.556, ranging from 0.374 to 0.714. Genetic distance of clusters ranged from 0.38 to 0.93 by unweighted pair-group method with arithmetical average based on Jaccard’s distance coefficients. These newly developed SSR markers indicate usefulness for variety identification related to seed dispute and distinctness, uniformity and stability (DUS) test for blueberry.

Ramie (Boehmeria nivea L.) is a hardy perennial herbaceous plant of the Urticaceae family and has been grown as a fiber crop in several countries including Korea for many centuries. Ramie leaves also have been traditionally used as a major ingredient of a type of rice cake called ‘Song-pyun’ in the Southwest area of Korea, especially Yeong-Gwang province. Despite its economic importance, the molecular genetics of ramie have not been studied in detail yet. Genetic resources of ramie were widely collected from domestic local sites by Bioenergy Crop Research Center (RDA) and Yeong-Gwang Agricultural Technology Center. For the systematic and efficient management of the genetic resources, we developed SSR (simple sequence repeat) markers of ramie. To do this, we generated microsatellite-enriched genomic DNA libraries using magnetic bead hybridization selection method. 247 non-redundant contigs containing SSR motif were generated using nucleotide sequences of 376 clones from the libraries. Primer sets were designed from the flanking sequences of the repeat motif. Finally, we selected 10 SSR markers, possibly showing polymorphism among the genetic resources. Results on the genotype analysis of the ramie genetic resources using the SSR markers will be presented.

This experiment was carried out to compare the morphological traits of Korean, Chinese, Japanese and Southeast Asian(SEA) soybeans from RDA genebank. Days to flowering were ranged from 51 to 125 days with an average of 75 days. Those of China were the shortest with an average of 58 days and those of SEA were the longest with an average of 99 days. Growth days were the shortest with 94 days from China, and longest with 188 days from Korea and SEA. The 100 seed weight of soybeans was ranged from 3.4g to 46.4g, with an average 22.2g. The 100 seed weight was the lightest with an average 11.8g from SEA and the heaviest with an average 24.6g from Korea. In growth habit, over 50% of being collected from Korea, Japan and China were erect type, but 94% from SEA were intermediate type. The highest percentage of seed coat color was yellow(66.1%), followed by yellowing green(10.0%). As a result of cotyledon color in 760 black seed was 76.1% with yellow, 23.9% with green. Green cotyledon was much more in Korea(38.6%) and Japan(33.3%) than other countries. One thousand seven accessions from Korea, Japan, China and SEA were analyzed using 7 SSR markers. One hundred eighty alleles were detected with a lowest 16 at the Satt537 and a highest 35 at the Satt390. The average polymorphism information content(PIC) was 0.68, the highest with 0.7 in Japan. Gene diversity was the highest with 0.73 in China and Japan, while the lowest in SEA with 0.68.

Polyembryony in many citrus varieties is an impediment in breeding because it makes hard to identify hybrids after crossbreeding. So, it has become imperative for developing efficient methods to distinguish zygotic seedling generated from polyembryonic seed depending on citrus variety. Simple sequence repeat(SSR) marker is one of useful systems for such purpose. However, SSR markers to separate zygotic seedlings derived from the crossbreeding between ‘Marita unshiu’ (Citrus unshiu) ‘Seongjeon’ and ‘Shiranuhi mandarin’ [(C. unshiu x C. sinensis) x C. reticulata] ‘Hallabong’ have not been developed yet. In this study we tried to identify an effective SSR marker to screen zygotic seedling after crossbreeding between ‘Seongjeon’ and ‘Hallabong’. For this investigation, 387 seedlings were generated from 114 seeds produced from crossing those two varieties. A total of 116 SSR markers were tested to identify a special marker for distinguishing origin, zygotic or nucellar seedling. As a result, two markers, SSR012 and SSR093, were found to be more effective than other markers. These two SSR markers might be useful to select zygotic individuals in crossbreeing between ‘Seongjeon’ and ‘Hallabong’.

In this study we evaluate the informative and efficiency of Simple Sequence Repeat (SSR) and Sequence Specific Amplified Polymorphism (SSAP) markers for genetic diversity, genetic relationship and population structure among 87 super sweet corn inbred lines generated by different origins. The SSR showed relatively higher level of the average gene diversity and shannon’s information index value than that of the SSAP. To assess genetic relationship and to characterize among 87 super sweet corn inbred lines using the SSR and SSAP markers. The dendrogram using SSR marker divided into nine groups of clusters were observed at the genetic similarity value 53.0%. For SSAP marker, Total three main clusters were confirmed in genetic similarity value at 50.8%. Result of combine data for SSR and SSAP markers showed six subgroup were detected in genetic similarity at 53.5%. To confirm population structure, the total 87 super sweet corn inbred lines were divided into groups I, II and admixed group based on membership probability 0.8 for SSR and SSAP markers. However population structure using combine data was K=3 and divided into group I, II, III and admixed group. This study has demonstrated the comparative analysis of SSR and SSAP for the study of genetic diversity and the genetic relationship for super sweet corn inbred lines. Thus, the results of this study will be useful to maize breeding programs in Korea.

In order to assess the genetic diversity, phylogenetic relationships and population structure of Korean rice landraces, 76 accessions were estimated using agronomical traits and SSR markers. Among 11 agronomical traits, amylose content (AC) was the trait with the largest variance with values ranged from 4.9 to 28.39 %, while grain length (GL) was the lowest variance ranged from 4.4 to 5.9 cm. In the result of PCA, the first PC with Eigen value of 217.5 explained 60.3% of the total variance. Culm length (CL) was the variable with the largest positive loadings. Growth period (GP) was the positive variances, while AC was the negative variance. The second PC with Eigen value of 80.6 explained an additional 22.4% of the total variance. Growth period (GP) was variable with highest positive loading. Amylose content (AC) was variable with high positive, while CL was the negative variance. The 49 SSR markers produced a total of 473 alleles with an average of 9.6 alleles. The polymorphism information content (PIC) was in the range 0.11 to 0.93. The observed heterozygosity ranged from 0.12 to 0.39, with an average of 0.61. 76 rice accessions showed two subpoplations and three groups based on SSR markers. Group I and Gropu II appertained Pop-2 and Pop-1 subpopulation, respectively. They showed similar agronomical traits. Group III consisted 7 rice accessions predominantly appertained to Pop-1. These results provide insight into the characters of Korean rice landraces and help to improve our knowledge of rice breeding

This study was conducted to construct a DNA marker database for 38 plum varieties collected in Korea using simple sequence repeat (SSR) markers. A set of 61 SSR primer pairs was tested to select polymorphic SSR markers between 8 varieties. Among the 61 primer pairs, 21 showed polymorphism, reproducibility and easy scoring. The genetic relationship between the 21 SSR markers and 38 varieties was analyzed. A total of 210 polymorphic amplified fragments were obtained with the 21 SSR markers. Three to seventeen SSR alleles were detected for each locus, with an average of 10.0 alleles per locus. Average polymorphism information content (PIC) was 0.758, with a range from 0.549 to 0.870. A total of 210 SSR marker loci were used to calculate Jaccard’s distance coefficients for cluster analysis by an unweighted pair-group method with arithmetical average (UPGMA). The genetic distance ranged from 0.06 to 1.00 in 38 varieties. Out of 38 plum varieties, 32 were identified using the 21 SSR markers. Therefore, these SSR markers may be employed to complement distinctness, uniformity, and stability (DUS) tests or as potential tools to solve seed disputes regarding plums.

본 연구는 12개의 색소 및 비색소옥수수 계통들에 대하여 300개의 SSR마커를 이용하여 유전적 다양성, 계통유연관계 및 집단구조 분석을 실시하였다. 유전적 다양성 분석 결과, 300개의 SSR primer들은 색소 및 비색소옥수수 12계통들에 서 총 1,331개의 대립단편을 증폭시켰으며, SSR primer당 대 립단편들은 최소 2개에서 최대 10개까지 나타나 평균 4.44개가 증폭되었다. MAF는 0.25에서 0.92의 범위로 나타났고, 평균값은 0.48을 나타내었다. 총 1,331개의 대립단편 중에서 221개의 대립단편들은 색소옥수수 계통들에서 특이적으로 나 타났고, 408개의 대립단편은 비색소옥수수 계통들에서 특이 적으로 나타났으며, 나머지 702개의 대립단편들은 색소 및 비색소 계통들에서 공통으로 확인되었다. 그리고 총 163개의 SSR 마커에서 색소옥수수 특이적 대립단편이 확인되었다. 집 단구조에 대한 분석 결과에서 12개의 색소 및 비색소옥수수 핵심집단 계통들은 groups I, II, III, admixed group으로 구 분되었다. 본 연구결과는 옥수수 육종연구에서 색소 및 비색 소옥수수 계통들의 식별에 대한 유용한 정보를 제공할 것이다

본 연구는 농업유전자원센터에 보존 되어있는 콩 유전자원 중 한국 재래종 자원 880점을 대상으로 농업적 특성을 조사하고, 분자마커를 이용하여 유전적 다양성을 분석하였으며 결 과는 다음과 같다. 1. 개화일수는 51∼104일, 평균 74.4일이었으며 성숙일수는 28∼106일, 평균 72.2일이었고 생육일수는 101∼188일, 평균 146.6일이었다. 지역별 평균 개화일수는 강원지역이 69.5일로 가장 짧고, 제주지역이 77.9일로 가장 길었다. 또한 평균 생육일수는 강원지역이 평균 140.6일로 가장 짧고, 제주지역이 152.8일로 가장 긴 것으로 나타났다. 대조품종의 생육일수는 태광콩 133일, 대원콩 143일, 일품검정콩 129일, 풍산나물콩 146일과 비교하였을 때 중만생종인 태광콩보다 생육일수가 길어 우리나라 재래종에 만생종이 많음을 알 수 있었다. 2. 100립중의 범위는 4.3∼46.4 g이며, 평균 26.1 g이고, 립중별 분포는 대립종의 비율이 39.2%로 가장 많고, 중립종 30.8%, 극대립종 17.5%, 소립종 8.8%, 극소립종 3.8% 순으로 중립종 이상의 비율이 87.5%로 대부분이었다. 3. 종피색은 검정색의 비율이 52.4%로 가장 많았고, 황색 28.5% 순이었다. 지역별로는 경남지역 수집자원은 황색종피 비율이 높고, 제주지역은 황색, 녹색, 검정색의 비율이 유사하였으며, 경기지역은 녹색종피의 비율이 28.8%로 황색종피보다 많았다. 4. 검정콩 460점을 대상으로 자엽색 조사 결과 황색이 59.7%, 녹색이 40.1%이었다. 자엽색의 지역별 분포는 매우 커서 제주와 경남지역은 녹색 자엽비율이 각각 83.3%와 62.7%로 황색 자엽보다 월등히 많았으며, 전북과 전남지역의 녹색자엽 비율은 각각 5.7%와 13.8%로 극히 낮아 지역별 차이를 보였다. 5. 기초특성 평가 자원중 350점에 대해 7개의 SSR 마커를 이용하여 프로파일링한 결과 총 110개의 대립인자(allele)가 확인되었으며 각 유전자좌별로 대립인자수는 10개에서 26개 평균 15.7개였다. 유전적 다양성(PIC)은 0.711로 비교적 높았으며, 각 유전자좌별 allele의 수는 경북지역이 9.7로 가장 많고, 유전적 다양성은 제주가 0.693으로 가장 높았다.

Seed & Variety Service for Plant Variety Protection (PVP). We previously constructed DNA profile database for identifying 159 lettuce varieties using 60 simple sequence repeat (SSR) markers. In this study, we selected optimum markers from previously applied markers and new SSR markers for the standardization of DNA profile database of 65 commercial lettuce varieties containing 18 PVP varieties distributed in Korea. Twenty-eight SSR markers from 60 SSR markers were selected for characterization with 65 commercial lettuce varieties according to the reproducibility, polymorphism, band pattern of marker and easiness of scoring. Out of 156 SSR primers, we additionally selected 11 new SSR primer pairs showed polymorphisms between 8 varieties and repetitive reproducibility on capillary electrophoresis system. Totally 127 polymorphic amplified fragments were obtained using 39 SSR markers. Two to seven SSR alleles were detected for each locus with an average of 3.3 alleles per locus. Average polymorphism information content was 0.517, ranging from 0.281 to 0.771. Genetic distance of clusters ranged from 0.29 to 0.96 by unweighted pair-group method with arithmetical average based on Jaccard’s distance coefficients. A total of 65 commercial lettuce varieties were discriminated by 39 SSR marker genotypes. These SSR profile database developed will be continually characterized for the standardization of DNA profiles for lettuce commercial varieties

Chrysanthemum (Dendranthema grandiflorum Ramat.) is a member of the Asteraceae and 588 varieties obtained Plant Variety Protection and 601 varieties were registered to Korea Seed & Variety Service for marketing. Thus, chrysanthemum is one of the important horticultural crop and having a possibility for infringement of Plant Breeder’s Rights. We investigated the genetic relationship of 20 chrysanthemum varieties using simple sequence repeat (SSR) markers. A total 335 SSR primer sets were screened for identification of 20 varieties. With 335 SSR primers, seventeen SSR primers showed polymorphism and reproducibility between 20 varieties. A total of 77 polymorphic fragments were identified by 17 SSR markers. Two to eight SSR alleles were detected for each SSR locus with an average of 4.53 alleles per locus. The polymorphism information content values ranged from 0.408 to 0.825 with an average of 0.45. Genetic distance of 20 varieties ranged from 0.36 to 0.73 by unweighted pair-group method with arithmetical average based on Jaccard’s distance coefficients. All of twenty varieties were distinguished by 17 marker genotypes. Future work will be investigated to construct DNA profile database for large number of chrysanthemum varieties using DNA sequencer.

Sorghum (Sorghum bicolor (L.) Moench) has been cultivated for cereal grain which has been traditionally used for steaming with rice in Korea. Various Korean sorghum varieties have been developed and distributed for farmers and consumers to meet their needs. Korean sorghum grains have been mostly sold at higher price in the market than sorghum grains imported from abroad. However, no varietal identification method was established to support fair trade in the cereal market. The objective of this study is to develop the identification method of Korean sorghum varieties using a multiplexed fingerprinting platform of SSR markers. One marker for the waxy allele and nine SSR markers were carefully selected based on their product sizes for the multiplexing. A robust multiplexed combination was revealed from serially designed experiments for the optimization of multiplex PCR. Five varieties and two elite breeding lines could be separated with their unique fingerprinting pattern from other sorghum individuals collected over the world. The platform separated most of individuals tested in this study, remaining three genotypes contained two or three identical individuals. The technique may be applied to detect closely-related individuals including full sibling progeny

본 연구는 강원도농업기술원 옥수수연구소에서 새로운 기능성 색소옥수수 품종을 개발하기 위해 육성한 총 12개 의 색소옥수수 계통들과 찰옥수수 및 일반옥수수 계통들에 대하여 SSR 분자마커를 이용하여 유전적 다양성, 집단 구조 및 association mapping 분석을 실시하였다. 그 결과 분석에 이용된 300개의 SSR primer들은 12개의 옥수수 자 식계통들에서 총 1,331개의 대립단편을 나타내었으며, 각 SSR primer 당 증폭된 대립단편의 수는 2개(umc1515, umc2249, umc1158, umc1659, phi102228, umc2262, umc1058, nc004, phi092, umc2308, umc1314, umc1178, umc1352a, umc2092, phi057, umc1139, umc1473, umc2338, umc2093, umc1107, umc1054)에서 10개(mmc0111)의 범위로 나타났 으며, 평균 대립단편 수는 4.44개였다. 집단구조 분석결과, 12개의 옥수수 자식계통들은 groups I, II, III, admixed group으로 구분되었다. 2개의 자식계통(10S4015, 10S4026)은 group I에 포함되었고, Group II는 총 4개의 자식계통 (Mo17, B14A, HW7, HW3)이 포함되었다. 그리고 4개의 자식계통(11CS4117, 11CS4124, 11CS7014, HW9)은 Group III에 포함되었으며, 2개의 자식계통(10S4032, KW7)은 admixed group에 포함되었다. 더욱이 본 연구에서는 색소 및 비색소옥수수 자식계통들에서 분석에 이용된 300개 SSR 마커와 4개의 양적 형질(간장, 착수고, 간경, 수술길이) 과의 연관성을 분석하기 위해서 population structure(Q) 값을 이용하여 Q GLM 분석을 실시하였다. 0.01의 유의수준 에서, Q GLM 분석을 이용하여 총 17개의 SSR 마커가 4개의 형질과 association을 확인하였다. 본 연구에서 12개의 색소 및 비색소옥수수 자식계통들에 대한 유전적 다양성, 집단구조 및 association mapping 분석의 결과는 앞으로 강원도농업기술원 옥수수연구소에서 기능성 색소옥수수 품종개발을 위한 계통 육성 및 교배조합 구성 등에 유용 한 정보를 제공할 것으로 기대한다.

Korea is a origin of three basic species, P. ussuriensis, P. pyrifolia and P. fauriei. Genetic relationship among Korean pear cultivars compared with their parents were also identified that they are closely related P. pyrifolia, P. ussuriensis and/or hybrid between two species. SSRs or Microsatellites are co-dominant and typically neutral inheritance showing high degree of polymorphism, large number of alleles per locus, abundance in genomes, and suitability for automation. SSR markers were developed in apple and pear where they were used for construction of genetic linkage maps, evaluation of the genetic diversity, cultivar identification, genotype identification, and in the determination of genetic relatedness. Many apple (Malus × domestica Borkh.) SSRs would be useful for genetic mapping in European and Asian pears in previous experiments and cross-species amplification was observed between apple and pear. The objectives of this study were to develop polymorphic SSR markers in ‘Whangkeumbae’ and ‘Minibae’, which were chosen as the representative cultivars of P. pyrifolia and P. ussuriensis in each among Korean pears, from ‘Golden Delicious’ genomic sequences generated by next generation sequencing technology and to evaluate the utility of the SSR markers based on ‘Golden Delicious’ sequences. Of 51 SSR markers, 18 were polymorphic in ‘Whangkeumbae’ and ‘Minibae’. The cross-species transportability of primers designed in ‘Golden Delicious’ sequences makes SSR markers more useful, given the current high level of investment in mapping the genomes of related Rosaceae.

Foxtail millet (Setaria italica L.) is the second most widely cultivated species of millet, especially in East Asia and is a tractable experimental model crop for studying functional genomics of millets. However, insufficient researches had been conducted about the foxtail millet germplasm and is significantly impeding its genetic improvement. We attempted to develop EST-derived-SSR (eSSR) markers and utilize them in genetic comparison of germplasm and transferability. A total of 66,027 foxtail millet EST sequences and 42,754 genomic sequence were deduced transcriptom. Approximately 42,000 single tone contigs were generated using DNAstar 5.0 software for redundancy minimization. Nearly 33% of the 14,012 unigenes contained SSRs, but primers were designed for a total of 314 microsatellites concentrating with more than 24 bp of repeats. A total of 314 primers were successfully designed with more than 24 bp of repeats. From these microsatellites, 56 primer pairs were showed polymorphism with over than 15 bp differences among 96 accessions collected from different countries. Polymorphic information content (PIC) value ranged from 0.020 to 0.700 with an average of 0.381 indicating moderate level of informativeness within these EST-SSRs markers. The EST-SSR markers developed in this study will serve as a useful source for genetic studies, such as genetic variability, transferability, association mapping, and molecular breeding