• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.57-57
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• 2017

Chrysanthemum is one of the most important floral crop in Korea produced about 7 billion dollars (1 billion for pot and 6 billion for cutting) in 2013. However, it is difficult to breed and to do genetic study because 1) it is highly self-incompatible, 2) it is outcrossing crop having heterozygotes, and 3) commercial cultvars are hexaploid (2n = 6x = 54). Although low-density genetic map and QTL study were reported, it is not enough to apply for the marker assisted selection and other genetic studies. Therefore, we are trying to make high-density genetic mapping using GBS with about 100 $F_1s$ of C. boreale that is oHohhfd diploid (2n = 2x = 18, about 2.8Gb) instead of commercial culitvars. Since Chrysanthemum is outcrossing, two-way pseudo-testcross model would be used to construct genetic map. Also, genotype-by-sequencing (GBS) would be utilized to generate sufficient number of markers and to maximize genomic representation in a cost effective manner. Those completed sequences would be analyzed with TASSEL-GBS pipeline. In order to reduce sequence error, only first 64 sequences, which have almost zero percent error, would be incorporated in the pipeline for the analysis. In addition, to reduce errors that is common in heterozygotes crops caused by low coverage, two rare cutters (NsiI and MseI) were used to increase sequence depth. Maskov algorithm would also used to deal with missing data. Further, sparsely placed markers on the physical map would be used as anchors to overcome problems caused by low coverage. For this purpose, were generated from transcriptome of Chrysanthemum using MISA program. Among those, 10 simple sequence repeat (SSR) markers, which are evenly distributed along each chromosome and polymorphic between two parents, would be selected.

• Journal of Microbiology and Biotechnology
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• 제20권4호
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• pp.683-692
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• 2010

Morphologically, nine different slow-growing protoclones were screened from regenerated protoplasts of heterokaryotic Agaricus bisporus. As such, the present study is the first report on differentiating homo- and heterokaryotic protoclones using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. Among 80 primers tested, the seven ISSR and seven RAPD primers selected for the analysis generated a total of 94 ISSR and 52 RAPD fragments, respectively. The ISSR fingerprinting also detected more polymorphic loci (38.29%) than the RAPD fingerprinting (34.61%). A principal coordinate analysis (PCA) was employed to evaluate the resolving power of the markers as regards differentiating protoclones. As a result, the mean polymorphism information content (PIC) for each marker system (i.e., 0.787 for RAPD and 0.916 for ISSR) suggested that ISSR is more effective for determining polymorphisms. The dendrograms constructed using RAPD, ISSR, and an integrated RAPD and ISSR marker system were highly correlated with one another as revealed by a high Mantel correlation (r= 0.98). The pairwise similarity index values also ranged from 0.64 to 0.95 (RAPD), 0.67 to 0.98 (ISSR), and 0.67 to 0.98 (RAPD and ISSR), whereas the mean similarity index values of 0.82, 0.81, and 0.84 were obtained for the RAPD, ISSR, and combined data, respectively. As there was a good correspondence between the RAPD and ISSR similarity matrices, ISSR would appear to be an effective alternative to RAPD in the genetic diversity assessment and accurate differentiation of homo- and heterokaryotic protoclones of A. bisporus.

• 생명과학회지
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• 제21권3호
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• pp.468-472
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• 2011

DNA 마커는 유전현상 분석이나 품종육성에 널리 사용되고 있다. 본 연구에서는 내냉성 오이 계통인 'NC76'과 냉해 감수성 계통인 'GY14'로부터 내냉성 연관 마커을 목적으로 기존의 총 995개 SSR 마커의 다형성을 평가하였다. Agarose gel 전기영동법으로 'NC76과 'GY14' 간 PCR증폭 산물의 길이 다형성을 보이는 145개 SSR 마커를 개발하였으며, high resolution melting (HRM) 기술을 사용하여 염기서열 다형성을 보이는 30개의 SSR 마커를 확인하였다. 개발된 175개 SSR 마커 중 20개 마커를 선발하여 'NC76'과 'GY14' 간 $F_2$ 분리 집단에 대한 연관지도를 작성하였으며 그 결과 13개의 마커가 예상했던 연관군에 일치하여 위치됨을 확인할 수 있었다. 따라서 본 연구에서 확인된 175개의 SSR 마커는 향 후 냉해 저항성 연관 마커 개발을 위한 오이 유전자 지도 작성 및 이를 통한 품종 육성에 크게 활용될 수 있을 것으로 기대된다.

• Journal of Plant Biotechnology
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• 제47권4호
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• pp.298-308
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• 2020

Genetic diversity among Thaumatococcus daniellii populations in the southwestern region of Nigeria were assessed using morphometric and molecular markers to determine the population structure and existing genetic relationship for its improvement, conservation and sustainable utilisation. Populations from five locations in each of the six states were used for the study. Morphometric data were collected on folia characters and analysed for variability. Genome DNA was isolated from the plant leaf and amplified by polymerase chain reaction with inter-simple sequence repeat markers (ISSR) to determine the allelic polymorphism, marker effectiveness and genetic relationship of the population. The results showed significant variations in petiole length and leaf dimensions of the populations within and across the states. These morphometric traits are the major parameters that delimit the populations and they correlated significantly at P≤0.05. Analysis of the electrophoregram showed that the ISSR markers are effective for the diversity study. A total of 136 loci were amplified with an average of 7.16 loci per marker, 63.2% of the loci were polymorphic. The Principal Coordinate Analysis revealed that seven factors accounted for 81.6% of the variation and the dendrogram separated the populations into two major groups at a genetic distance of 10 (about 90% similarity) with sub-groups and clusters. Most populations within the state had a high degree of similarity, nonetheless, strong genetic relationship exists among populations from different states. The close relationship between populations across the states suggests a common progenitor, which are likely separated by ecological or geographical isolation mechanisms.

• Journal of Plant Biotechnology
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• 제35권4호
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• pp.257-263
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• 2008

Improvement of the macro- and micro-elements density of rice (Oryza sativa L.) is gradually becoming a new breeding objective. In this study, the genomic regions associated with potassium, calcium, magnesium and iron content in rice grain were identified and characterized by using a doubled haploid (DH) population. Fifty-six simple sequence repeat (SSR) and one hundred and twelve sequence tagged site (STS) markers were selected to construct the genetic linkage map of the DH population with a full length of 1808.3cM scanning 12 rice chromosomes. Quantitative trait loci (QTLs) were detected, and QTL effects and QTL interactions were calculated for five traits related to macro- and micro-elements in the DH population from a cross between 'Samgang' (Tongil) and 'Nagdong' (Japonica). Twelve QTLs were located on five chromosomes, consisting of two QTLs for potassium, three QTLs for calcium, two QTLs for magnesium, one QTL for iron content and four QTLs for the ratio of magnesium to potassium (Mg/K). Among them, qca1.1 was detected on chromosome 1 with an LOD value of 8.58 for calcium content. It explained 27% of phenotype variations with increasing effects from 'Samgang' allele. Furthermore, fifteen epistatic combinations with significant interactions were observed on ten chromosomes for five traits, which totally accounted for 4.19% to 12.72% of phenotype variations. The screening of relatively accurate QTLs will contribute to increase the efficiency of marker-assisted selection (MAS), and to accelerate the establishment of near-isogenic lines (NILs) and QTL pyramiding.

• The Plant Pathology Journal
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• 제31권4호
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• pp.402-413
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• 2015

Yellow rust (stripe rust), caused by Puccinia striiformis f. sp. tritici, is one of the most destructive foliar diseases of wheat in Egypt and worldwide. In order to identify wheat genotypes resistant to yellow rust and develop molecular markers associated with the resistance, fifty F8 recombinant inbred lines (RILs) derived from a cross between resistant and susceptible bread wheat landraces were obtained. Artificial infection of Puccinia striiformis was performed under greenhouse conditions during two growing seasons and relative resistance index (RRI) was calculated. Two Egyptian bread wheat cultivars i.e. Giza-168 (resistant) and Sakha-69 (susceptible) were also evaluated. RRI values of two-year trial showed that 10 RILs responded with RRI value >6 <9 with an average of 7.29, which exceeded the Egyptian bread wheat cultivar Giza-168 (5.58). Thirty three RILs were included among the acceptable range having RRI value >2 <6. However, only 7 RILs showed RRI value <2. Five RILs expressed hypersensitive type of resistance (R) against the pathogen and showed the lowest Average Coefficient of Infection (ACI). Bulked segregant analysis (BSA) with eight simple sequence repeat (SSR), eight sequence-related amplified polymorphism (SRAP) and sixteen random amplified polymorphic DNA (RAPD) markers revealed that three SSR, three SRAP and six RAPD markers were found to be associated with the resistance to yellow rust. However, further molecular analyses would be performed to confirm markers associated with the resistance and suitable for marker-assisted selection. Resistant RILs identified in the study could be efficiently used to improve the resistance to yellow rust in wheat.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 춘계학술대회
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• pp.45-45
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• 2020

A large number of expressed sequence tags (ESTs) in public databases have provided an opportunity for the systematic development of simple sequence repeat (SSR) markers. EST-SSRs derived from conserved coding sequences show considerable cross-species transferability in related species. In the present study, we assessed the utility of foxtail millet EST-SSRs in barnyard millet. A total of 312 EST-SSRs of foxtail millet were tested using 84 Echinochloa crus-galli germplasm accessions; a high rate of transferability (62%) and 46 primer sets (13%) were shown the polymorphism in barnyard millet. The 13% of functional EST-SSRs) was demonstrated between cereals and barnyard millet. SSR marker profile data were scored for the computation of pairwise distances as well as a Neighbor Joining (NJ) tree of all the genotypes. The averaged values of gene diversity (H_E) and polymorphism information content (PIC) were 0.213 and 0.179 within populations, respectively. The 84 barnyard millet germplasm accessions were divided into five different groups, which agreed well with their geographical origins. The exotic 12 accessions of India type barnyard millet (E. frumentacea) were all separated form Korean local collection genotype. The present results provide evidence of divergence between cultured and wild type barnyard, as a millet and grass. The polymorphic SSR markers indicated in this study were of great value in analysis of genetic diversity that can be further used for crop improvement through breeding.

• 원예과학기술지
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• 제31권3호
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• pp.337-343
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• 2013

본 연구의 목적은 SSR과 SRAP 마커 시스템을 이용하여 팔레놉시스 14품종 간 유전적 거리를 비교하고, SSR 마커를 이용하여 품종 간 구분을 하기 위한 것이다. 전체적으로 111개의 SSR 프라이머와 30조합의 SRAP primer를 먼저 스크리닝하였다. 국립원예특작과학원에서 보존중인 국내 육성품종을 포함한 14품종의 팔레놉시스에서 12개의 SSR 프라이머와 30조합의 SRAP 프라이머에서 높은 다형성을 보였다. 증폭된 DNA 단편들은 acrylamide gel에서 분리시킨 후 silver staining 방법으로 검출하였다. SSR 마커 55개와 SRAP 419개로, 총 474개의 마커를 획득하였으며 이를 유전적 다양성 분석에 사용하였다. 다형성 밴드들은 MVSP 3.1프로그램을 이용하여 유전적 유사도와 UPGMA clustering 분석을 위해 scoring 되었다. 14 팔레놉시스 품종은 SRAP과 SSR 분석을 통해 각각 0.683과 0.66의 유사도 지수에서 3그룹으로 분류되었다. 또한 SSR 20번과 22번만으로도 이들 육성 품종을 구분할 수 있었다. 이 결과는, SSR 분석은 팔레놉시스 품종간 구분에 효과적이고 SRAP은 염기서열의 정보가 없을 때 유전적 다양성 분석에 유용하다는 것을 보여준다. 이번 연구된 SSR과 SRAP 마커들은 팔레놉시스의 유전자형 판별, 유전자원 보존, 유전적 근연관계를 분석하는데 유용한 기술이 될 것이다.

• 한국해양생명과학회지
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• 제6권1호
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• pp.38-46
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• 2021

꼬막 종류 중 하나인 Tegillarca granosa는 해양 이매패류로서 한국, 중국, 일본 등의 중요한 수산 자원 중 하나이다. 꼬막의 염색체 수는 2n=38로 알려져 있지만, 유전체의 크기와 유전 정보에 대해서는 아직 명확하게 알려져 있지 않다. 꼬막의 유전체 크기 예측을 위하여 NGS Illumina HiSeq 플랫폼을 이용하여 얻은 짧은 DNA 서열 정보를 통하여 in silico 분석으로 유전체 크기를 분석하였다. 그 결과 꼬막의 유전체 크기는 770.61 Mb로 예측되었다. 이후 MaSuRCA assembler를 통하여 드래프트 게놈 조립 작업을 수행하고, QDD pipeline을 이용하여 SSR (simple sequence repeats) 분석을 수행하였다. 꼬막의 유전체로부터 43,944개의 SSR을 발굴하였으며, 다이-뉴클레오타이드(di-nucleotide) 69.51%, 트라이-뉴클레오타이드(tri-nucleotide) 16.68%, 테트라-뉴클레오타이드(tetra-nucleotide) 12.96%, 펜타-뉴클레오타이드(penta-nucleotide) 0.82% 그리고 헥사-뉴클레오타이드(hexa-nucleotide) 0.03%로 구성되었다. 이후 꼬막의 유전적 다양성 연구에 활용할 수 있는 100개의 마이크로새틀라이트 마커의 프라이머 세트를 선별하였다. 앞으로 이번 연구를 통해서, 꼬막의 집단유전학적 연구와 유전적 다양성을 규명하는데 도움이 될 것이며, 나아가 동종들 간의 원산지 분류를 알아낼 수 있을 것이다.

• 한국약용작물학회지
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• 제20권4호
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• pp.277-285
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• 2012

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.