본 연구는 ICuGI Database를 활용하여 수박 상용 F1 품종의 순도검정용 EST-SSR 마커를 개발하기 위해 수행되었다. 총 353개 EST-SSR primer set을 선발하여 (주)NH종묘의 수박 F1 품종 7종과 각 품 종의 양친 11종에 대해 검정하였다. 이중 1차 테스트한 96개 primer set 중, ‘오렌지’는 primer WMU0056, ‘흑보’는 WMU0400, ‘신동’은 WMU0056와 WMU0400, ‘새로나’는 WMU0056, WMU0400, WMU0529에 대해 각 품종의 양친들이 다형성이 보였고 F1 개체에서는 이형접합의 유전자형 을 보였다. ‘해동’ F1의 순도검정용 마커를 찾기 위해 추가적인 122개의 primer set에 대해 PCR을 수 행한 결과, WMU0056, WMU0400, WMU0580, WMU1211, WMU4136, WMU448이 순도검정에 적합한 것으로 나타나, WMU0056와 WMU0400이 ‘해동’에서도 유용할 수 있었다. ‘꿀나라’와 ‘황피’의 경우에는 공시된 타 품종들에 비해 양친간 다형성율이 각각 5%와 2%로 매우 낮아 모든 353개 primer set을 테스트하였으며, 그 결과 ‘꿀나라’는 WMU5339, ‘황피’는 WMU7003이 순도검정 마커로 적합 한 것으로 확인되었다. 개발된 마커를 이용하여 실제 농가채종된 4개의 F1 품종의 순도를 검정한 결과, 모두 97.5% 이상의 순도로 확인되었다. 이와 같이 ICuGI Database에 공시된 수박 EST-SSR 마커는 공 우성의 유전자 특이적 마커로서 F1 순도검정에 효과적으로 사용될 수 있었다.

우리나라 참나리 2배체와 3배체 중 임의 선발된 56 개 지역의 참나리에 대하여 EST-SSR이용하여 각 genome 간의 유전적 변이와 유연관계를 분석하고자 수행되었다. 최근 백합속에서 개발된 19개의 EST-SSR 중에서 7개의 primer가 참나리 2, 3배체 종내 유전적 변이 분석에 적합한 것으로 나타났다. 서해안, 남해안 제주도의 원거리 지역에 분포하는 2배체 참나리들은 지역에 관계없이 다양한 유전적 변이를 나타내었으나, 소청도, 울릉도 및 내륙에 분포하는 3배체 참나리는 비교적 단순한 변이를 나타내었다. 다형성을 나타낸 총 121개의 SSR 밴드를 사용하여 UPGMA 방법으로 계 통도를 작성한 결과 2배체 집단에서는 유전적 변이가 다양한 반면 3배체 집단에서는 비교적 변이가 적은 것 으로 나타났다. 2배체 집단에서 아차도 계통들이 남해 안의 다른 2배체와 뚜렷이 구분되는 cluster를 형성하 였고, 3배체 집단에서는 소청도 참나리가 다른 지역과 는 구별되는 독특한 밴드패턴을 나타내었다. 본 연구에 서 선발된 EST-SSR마커들은 금후 참나리 2, 3배체 집단의 유연관계를 분석하는데 유용하게 사용될 수 있 을 것으로 생각된다.

In crop breeding program, information about genetic dissimilarity on breeding resources is very important to corroborate genealogical relationships and to predict the most heterozygotic hybrid combinations and inbred breeding. This study aimed to evaluate the genetic variation in Kenyan sunflower breeding lines based on simple sequence repeat (SSR). A total of 83 alleles were detected at 32 SSR loci. The allele number per locus ranged from 2 to 7 with an average of 2.7 alleles per locus detected from the 24 sunflower accessions and the average value of polymorphic information contents (PIC) were 0.384. A cluster analysis based on the genetic similarity coefficients was conducted and the 24 sunflower breeding resources were classified into three groups. The principal coordinates (PCoA) revealed 34% and 13.38% respectively, and 47.38% of total variation. It was found that the genetic diversity within the Kenyan sunflower breeding resources was narrower than that in other sunflower germplasm resources, suggesting the importance and feasibility of introducing elite genotypes from different origins for selection of breeding lines with broader genetic base in Kenyan sunflower breeding program.

Background : Codonopsis lanceolata is a perennial plant of Campanulaceae and mainly distributed in East Asia such as Korea, China, and Japan. C. lanceolata has a unique taste and aroma, and it is rich in minerals such as phosphorus and calcium, and vitamin B1 and B2, so our ancestors used the plant as medicinal herb and edible vegetable. However, systematic cultivation and development of varieties have not been achieved compared to demand or high added value. The genetic diversity and relationship analysis of the plants help to increase the efficiency of breeding through genetic variation. Methods and Results : Ten species of Codonopsis plants were used as materials and DNA was extracted from each 4 individuals per species and quantified at a concentration of 10 ng /㎕. The extracted DNA was pooled by species and PCR was performed using the EST-SSR marker developed based on C. lanceolata in the previous study. PCR amplification was carried out using a denaturation at 94℃ for 30 sec, annealing at 58℃ for 30 sec and extension at 72℃ for 30 sec, repeated for 35 total cycles. The PCR products were separated in a 4% agarose gell at 100 V for 40 min. Conclusion : In this study, C. lanceolata collections was determined among several Codonopsis species using these molecular marker. It is expected that the data of this study can be used as reference for genetic polymorphism analysis and related gene studies of Codonopsis species.

Understanding the genetic variation among landrace collections is important for crop improvement and utilization of valuable genetic resources. The present study was carried out to analyse the genetic diversity and associated population structure of 621 foxtail millet accessions of Korean landraces using 22 EST-SSR markers. A total of 121 alleles were detected from all accessions with an average of 5.5 alleles per microsatellite locus. The average values of gene diversity, polymorphism information content, and expected heterozygosity were 0.518, 0.594, and 0.034, respectively. Following the unweighted neighbor-joining method with arithmetic mean based clustering using binary data of polymorphic markers, the genotypes were grouped into 3 clusters, and population structure analysis also separated into 3 populations. Principal coordinate analysis (PCoA) explained a variation of 13.88% and 10.99% by first and second coordinates, respectively. However, in PCoA analysis, clear population-level clusters could not be found. This pattern of distribution might be the result of gene flow via germplasm exchanges in nearby regions. The results indicate that these Korean landraces of foxtail millet exhibit a moderate level of diversity. This study demonstrated that molecular marker strategies could contribute to a better understanding of the genetic structure in foxtail millet germplasm, and provides potentially useful information for developing conservation and breeding strategies.

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

Toxic (high phorbol esters) and nontoxic (low phorbol esters) jatropha accessions cannot be distinguished morphologically. Their seeds must be chemically analyzed through a complex and costly process using HPLC method. EST-SSR markers can be used to classify jatropha accessions with high and low phorbol esters. In this study, ninety-seven EST-SSR markers amplified the genomic DNA and showed polymorphism among 5 high phorbol esters accessions and 10 low phorbol esters accessions. These markers can be further exploited for jatropha improvement through marker-assisted breeding.

본 연구는 감 수집종의 분류 및 품종 육종을 위하여 EST-SSR 마커를 개발해 유전적 유연관계를 분석하고, 형태적 유연관계를 비교 분석하여 DNA 마커의 효율성을 극대화한 연구 결과이다. 경북농업기술원 상주감시험장에서 수집한 42품종을 대상으로 6가지의 양적형질(과실크기, 과고, 과경, 과경굵기, 과경길이, 종자크기)과 19가지의 질적형질(횡단면, 종단면, 골의 정도, 얕은 동심원 균열, 옆모양, 정부열과, 세로홈, 꽃받침 끝 주름, 배꼽 홈길이, 꽃받침 쪽의 홈, 꽃받침 크기)을 사용하여 형태적 유연관계를 분석하였다. 유전적 유연관계를 분석하기 위해 수집한 감에서 cDNA library를 만들어 sequence를 분석한 후, PCR을 통해 얻은 polymorphism이 인정되는 25개의 primer set에서 16개의 EST-SSR primer set를 선발하였다. 수집한 감 42품종의 형태적 유연관계와 개발한 14개의 EST-SSR 마커를 이용하여 유전적인 유연관계를 분석한 결과 형태적 유연관계에서는 여러 그룹이 형성되었지만 coefficient가 0.02 이하로 형성되어 형태적 특성을 사용해 분류하기는 어려웠다. 유전적 유연관계는 coefficient 0.77에서 3개 그룹으로 분류되어, 상주수수감과 상주수꽃감, 밀양반시와 밀양고동시, 영동반시와 영동수시는 각각 같은 그룹으로 분류되었다. 형태적 분석과 유전적 분석의 상관관계를 조사한 결과 형태적 분석의 유사도 거리와 유전적 분석의 유사도 거리 간의 값이 -0.03으로 유의성이 매우 낮게 나왔다. 본 실험에서 얻어진 분자마커는 (EST-SSR 마커) 국내 감육종 효율 증진뿐만 아니라 우수형질을 도입하는데 유용하게 이용할 수 있을 것으로 기대된다.

Genome duplication is an abundance phenomenon and in plant kingdom and consequently formed paralogous region. Korean ginseng (Panax ginseng C.A. Meyer) has a possibility of tetraploid by comparing chromosome numbers of relative species. During development of EST-SSR markers in Korean ginseng, most of primer sets have produced multiple bands in gel electrophoresis. In this study, for identifying origin of multiple bands, five EST-SSR markers showing multi-band were selected and two bands around expected size were sequenced. Sequence comparison classified the multiple bands into individual loci. Two bands can be identified by SNP or InDel variation with number of SSR units. Sequencing result represented that paralogous loci with high similarity were existence caused by recent duplication. One clear band were amplified with newly designed locus specific primer picked from SNP variation. SNP and InDel polymorphism between paralgous loci were useful for identifying each locus. This study will provide better understanding of ginseng genome and will be helpful for development of DNA markers.

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.

This study was carried out to develop expressed sequence tag-Simple sequence repeat (EST-SSR) markers of brown plant hopper resistance gene originated from a rice cultivar ‘Cheongcheong’ and sensitive rice cultivar ‘Nakdong’. Total RNA extracted from the leaves of ‘Cheongcheong’ and ‘Nakdong’ were used to synthesize a cDNA library. As a result of analyzing the cDNA library, EST-SSR sites were found and the EST-SSR primer sets were developed. This study enables to provide effective marker assisted selection (MAS) methods on the selection of white-backed planthopper resistance gene originated from a rice cultivar more simply, quickly and precisely. Furthermore, using this marker’s advantage of deriving from cDNA, it is possible to identity the white-backed planthopper resistance gene.