• Title/Summary/Keyword: SSR markers

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Genotyping of avian pathogenic Escherichia coli by DNA fragment analysis for the differences in simple sequence repeats

  • Han, Mi Na;Byeon, Hyeon Seop;Han, Seong Tae;Jang, Rae Hoon;Kim, Chang Seop;Choi, Seok Hwa
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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    • pp.257-262
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    • 2018
  • Avian pathogenic E. coli (APEC) causes severe economic losses in the poultry farms, due to systemic infections leading to lethal colisepticemia. It causes a variety of diseases from air sac infection to systemic spread leading to septicemia. Secondary infection contains opportunistic infections due to immunosuppression disease. Collibacillosis causes the great problems in the poultry industry in Korea. Thus, it is necessary to identify and classify the characteristics of E. coli isolate of chicken origin to confirm the diversity of symptoms and whether they are transmitted among the farms. Fragment analysis is identify the difference in the number of Variable-Number Tandem-Repeats (VNTRs) for genotyping. VNTRs have repeating structure (Microsatellite, Short tandem repeats; STR, Simple sequence repeats; SSR) in the chromosome. This region can be used as a genetic marker because of its high mutation rate. And various lengths of the amplified DNA fragment cause the difference in the number of repetition of the DNA specific site. The number of repetition sequences indicates the separated size of fragments, so the each fragments can be distinguished by specific samples. The results of the sample show that there is no difference in six microsatellite loci (yjiD, aidB, molR_1, ftsZ, b1668, yibA). There are differences among the farms in relation of the number of repetitions of other six microsatellite loci (ycgW, yaiN, yiaB, mhpR, b0829, caiF). Four (ycgW, yiaB, b0829, caiF) of these six microsatellite loci show statistically significant differences (P<0.05). It means that the analysis using four microsatellite loci including ycgW, yiaB, b0829, and caiF can confirm among the farms. Five E. coli samples in one farm have same SSR repetition at all markers. But, there are significant differences from other farms at Four (ycgW, yiaB, b0829, caiF) microsatellite loci. These results emphasize again that the four microsatellite loci makes a difference in the amplified DNA fragments, enabling it to be used for E. coli genotyping.

Genetic diversity analysis of tea (Camellia sinensis (L.) O. Kuntze) germplasm in Korea genebank

  • Hyeon, Do-Yun;Lee, Jeong-Ro;Jo, Gyu-Taek;Raveendar, Sebastin;Sin, Myeong-Jae;Lee, Gyeong-Jun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.63-63
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    • 2019
  • Tea plant (Camellia sinensis (L.) O. Kuntze) has been cultivated widely in many developing Asian, African, and South American countries, where it is the most widely consumed beverage in the world next to water. It has critical importance to understand the genetic diversity and population sturcutre for effective collection, conservation, and utilization of tea germplasm. In this study, 410 tea accessions collected from South Korea were analyzed using 21 SSR markers. Among 410 tea accessions, 85.4% (310 accessions) accessions were collected from Jeollanam-do. A total of 286 alleles were observed, and the genetic diversity and evenness were estimated to be averagely 0.79 and 0.61, respectively, across all the tested samples. Using discriminant analysis of principal components, the four clusters were detected in 410 tea accessions. Among them, cluster 1 showed higher frequency of rare alleles (less than 1%) than other clusters. Using calculation of the index of association and rbaD value, each cluster showed a clonal mode of reproduction. The result of AMOVA showed that most of the variation observed was within populations (99%) rather than among populations (1%). Our results might contribute to provide data about genetic diversity for the conservation of tea germplasm and for future breeding programs.

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Evaluation of Morphological Traits and Genetic Composition in Melon Germplasm (멜론 유전자원의 형태적 특성 및 유전적 구성 평가)

  • Lee, Seungbum;Jang, Ik;Hyun, Do Yoon;Lee, Jung-Ro;Kim, Seong-Hoon;Yoo, Eunae;Lee, Sookyeong;Cho, Gyu-Taek;Lee, Kyung Jun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.4
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    • pp.485-495
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    • 2020
  • Melon (Cucumis melo L.), one of the most important fruit crop species, is cultivated worldwide. In this study, a total of 206 melon accessions conserved at the National Agrobiodiversity Center (NAC) in RDA were characterized for nine morphological characteristics according to the NAC descriptor list. In addition, to confirm the genetic composition of each melon accession, genetic profiling was performed using 20 SSR markers. Among the 206 melon accessions, 159 (77.2%) were collected from Asia. The color of fruit flesh and skin were mostly 'white' (56.0%) and 'green' (49%), respectively. Days to female flowering (FD) and maturity (MD) of the accessions ranged from 58 to 72 and 17 to 63, respectively. The fruit length and width of the accessions ranged from 6.0 to 29.3 and 3.6 to 17.2 cm, respectively. The sugar content (SU) ranged from 2.5% to 13.2% with an average of 7.0%. In correlation analysis, SU showed positive and negative correlations with MD and FD, respectively. The accessions were classified into four clusters by cluster analysis. From the results of genetic profiling using 20 SSR markers, three accessions (K189118, K100486, and K190292) were expected to be inbred lines among 206 melon accessions. These results could expand the knowledge of the melon germplasm, providing valuable material for the development of new melon varieties to suit consumer tastes.

Identification of DNA Markers Related to Resistance to Herbicide Containing Mesotrione in Tongil Type Rice (통일형 벼에서 메소트리온계 제초제 저항성 연관 DNA marker 탐색)

  • Lee, Ji-Yoon;Cho, Jun-Hyeon;Lee, Jong-Hee;Cho, Su-Min;Kwon, Young-Ho;Park, Dong-Soo;Song, You-Chun;Ko, Jong-Min
    • Korean Journal of Breeding Science
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    • v.50 no.4
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    • pp.387-395
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    • 2018
  • This study was conducted to identify DNA markers related to resistance to herbicide containing mesotrione in Tongil type rice. Two Tongil type elite lines; Milyang154 and Suweon382, showed resistance to mesotrione, whereas the others were susceptible at 20 days after mesotrione application, and severe growth inhibition was observed in the remaining 13 lines. As a result of analysis of mesotrione resistance using 190 $F_2$ populations derived from a cross of Hanareum2 (susceptible) and Milyang154 (resistant), the mesotrione resistance locus was shown to be a single dominant gene with a 3:1 segregation ratio ($X^2=1.19$, P=0.31). To identify a DNA marker closely linked to the mesotrione resistance gene, bulked segregant analysis (BSA) was adopted. The DNA marker RM3501 was identified on chromosome 2 with a recombinant value of 0.53 to the mesotrione resistance gene. Mst1(t) was located between SSR (simple sequence repeat) markers RM3501 and RM324 with a physical map distance of 10.2 Mb-11.4 Mb on chromosome 2. The band pattern of agarose gel electrophoresis of the SSR marker RM3501 showed the same segregation pattern with respect to mesotrione treatment in 20 Tongil type varieties and a $BC_2F_2$ segregation population derived from a cross between Unkwang (resistant) and Hanareum2 (susceptible). Thus, the RM3501 DNA marker could be used in breeding programs for Marker Assisted Selection in mesotrione resistant rice breeding.

Current status and prospects of kiwifruit (Actinidia chinensis) genomics (참다래 유전체 연구 동향)

  • Kim, Seong-Cheol;Kim, Ho Bang;Joa, Jae-Ho;Song, Kwan Jeong
    • Journal of Plant Biotechnology
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    • v.42 no.4
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    • pp.342-349
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    • 2015
  • Kiwifruit is a new fruit crop that was commercialized in the late 1970s. Recently, its cultivation and consumption have increased rapidly worldwide. Kiwifruit is a dioecious, deciduous, and climbing plant having fruit with hairs and various flesh colors and a variation in ploidy level; however, the industry consists of very simple cultivars or genotypes. The need for efficient cultivar improvement together with the evolutional and biological perspectives based on unique plant characteristics, have recently encouraged genome analysis and bioinformatics application. The draft genome sequence and chloroplast genome sequence of kiwifruit were released in 2013 and 2015, respectively; and gene annotation has been in progress. Recently, transcriptome analysis has shifted from previous ESTs analysis to the RNA-seq platform for intensive exploration of controlled genetic expression and gene discovery involved in fruit ascorbic acid biosynthesis, flesh coloration, maturation, and vine bacterial canker tolerance. For improving conventional breeding efficiency, molecular marker development and genetic linkage map construction have advanced from basic approaches using RFLP, RAPD, and AFLP to the development of NGS-based SSR and SNP markers linked to agronomically important traits and the construction of highly saturated linkage maps. However, genome and transcriptome studies have been limited in Korea. In the near future, kiwifruit genome and transcriptome studies are expected to translate to the practical application of molecular breeding.

Locating QTLs controlling overwintering seedling rate in perennial glutinous rice 89-1 (Oryza sativa L.)

  • Deng, Xiaoshu;Gan, Lu;Liu, Yan;Luo, Ancai;Jin, Liang;Chen, Jiao;Tang, Ruyu;Lei, Lixia;Tang, Jianghong;Zhang, Jiani;Zhao, Zhengwu
    • Genes and Genomics
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    • v.40 no.12
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    • pp.1351-1361
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    • 2018
  • A new cold tolerant germplasm resource named glutinous rice 89-1 (Gr89-1, Oryza sativa L.) can overwinter using axillary buds, with these buds being ratooned the following year. The overwintering seedling rate (OSR) is an important factor for evaluating cold tolerance. Many quantitative trait loci (QTLs) controlling cold tolerance at different growth stages in rice have been identified, with some of these QTLs being successfully cloned. However, no QTLs conferring to the OSR trait have been located in the perennial O. sativa L. To identify QTLs associated with OSR and to evaluate cold tolerance. 286 $F_{12}$ recombinant inbred lines (RILs) derived from a cross between the cold tolerant variety Gr89-1 and cold sensitive variety Shuhui527 (SH527) were used. A total of 198 polymorphic simple sequence repeat (SSR) markers that were distributed uniformly on 12 chromosomes were used to construct the linkage map. The gene ontology (GO) annotation of the major QTL was performed through the rice genome annotation project system. Three main-effect QTLs (qOSR2, qOSR3, and qOSR8) were detected and mapped on chromosomes 2, 3, and 8, respectively. These QTLs were located in the interval of RM14208 (35,160,202 base pairs (bp))-RM208 (35,520,147 bp), RM218 (8,375,236 bp)-RM232 (9,755,778 bp), and RM5891 (24,626,930 bp)-RM23608 (25,355,519 bp), and explained 19.6%, 9.3%, and 11.8% of the phenotypic variations, respectively. The qOSR2 QTL displayed the largest effect, with a logarithm of odds score (LOD) of 5.5. A total of 47 candidate genes on the qOSR2 locus were associated with 219 GO terms. Among these candidate genes, 11 were related to cell membrane, 7 were associated with cold stress, and 3 were involved in response to stress and biotic stimulus. OsPIP1;3 was the only one candidate gene related to stress, biotic stimulus, cold stress, and encoding a cell membrane protein. After QTL mapping, a total of three main-effect QTLs-qOSR2, qOSR3, and qOSR8-were detected on chromosomes 2, 3, and 8, respectively. Among these, qOSR2 explained the highest phenotypic variance. All the QTLs elite traits come from the cold resistance parent Gr89-1. OsPIP1;3 might be a candidate gene of qOSR2.

Mapping QTLs for Agronomic Traits Using an Introgressin Line Population from a Cross between Ilpumbyeo and Moroberekan in Rice (일품벼/모로베레칸 이입계통을 이용한 농업형질 관련 QTL 분석)

  • Ju, Hong-Guang;Kim, Dong-Min;Kang, Ju-Won;Kim, Myung-Ki;Kim, Yeon-Gyu;Ahn, Sang-Nag
    • Korean Journal of Breeding Science
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    • v.40 no.4
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    • pp.414-421
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    • 2008
  • We conducted a QTL analysis of agronomic traits using 117 $BC_3F_5$ and $BC_3F_6$ lines developed from a cross between Ilpumbyeo and Moroberekan. Genotypes of 117 $BC_3F_5$ lines were determined using 134 simple sequence repeat (SSR) markers. A total of 832 Moroberekan chromosome segments with 410 homozygous and 422 heterozygous, respectively, were detected, and the genetic distance of introgression segments ranged from 0.5 cm to 112.1 cm. A linkage map constructed using 134 SSR markers was employed to characterize quantitative trait loci (QTL). The 117 $BC_3F_5$ and $BC_3F_6$ lines were evaluated for seven agronomic traits at two locations in 2006 and 2007 and at one location in 2007. A total of 26 QTLs were identified for seven traits including days to heading, and the phenotypic variance explained by each QTL ranged from 9.2% to 24.2%. Moroberekan alleles contributed positive effects in the Ilpumbyeo background at eleven QTL loci including panicle length and spikelets per panicle. Five QTLs, two for days to heading and one each for culm length, panicle length and spikelets per panicle were consistently detected in every occasions indicating that these QTLs are stable. Among them, two QTLs, spp6 for spikelets per panicle and pl6 for paniclel length were localized in the similar region. Increase in spikelets per panicle at this locus might be due to the increase in panicle length, because both traits were associated with increase in spikelets per panicle and panicle length due to the presence of the Moroberekan allele. These Moroberekan QTLs might be useful in breeding programs to develop high-yielding cultivars.

Identification of Quantitative Trait Loci Associated with Leaf Length. Width and Length/width Ratio in Two Recombinant Inbred Lines of Soybean (Glycine max L.) (두 집단의 재조합 근친교잡 계통 (RIL) 콩에서 엽장과 엽폭 및 장폭비와 관련된 양적헝질 유전자좌 분석)

  • Kim, Hyeun-Kyeung;Kang, Sung-Taeg
    • Journal of Life Science
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    • v.14 no.5
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    • pp.821-828
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    • 2004
  • The increasing apparent photosynthetic rate per leaf area may improve seed yield in soybean. Leaf area, length and width are related to the photosynthetic capability of the plant. In this study, two populations derived from the cross of Keunolkong, Shinpaldalkong and Iksanl0 were evaluated with simple sequence repeat (SSR) markers to identify length, width and length/width ratio of leaf. Leaf length/width ratio were significantly negative correlation with leaf width in K/S and K/I populations. In the K/S population, two minor QTLs for leaf length (LL) were found on LG Dlb+W and 1. Two QTLs on LG J and L were related to LL in K/I population. Two and three minor QTLs were identified in leaf width with total phenotypic variation of 13% and 18.04 in K/S and K/I populations, respectively. The leaf length/width ratio, two QTLs on LG I and L, and three QTLs on LG Cl, E and L were related to K/S and K/I populations, respectively. Thus it is assumed that the leaf traits are very much dependent on the genotype used and different breeding approach should be considered for the selection of favorite leaf traits in soybean breeding programs.

Genetic Diversity and Spatial Structure in Populations of Abelia tyaihyoni (줄댕강나무 (Abelia tyaihyoni) 집단의 유전다양성 및 공간구조)

  • Jeong, Ji-Hee;Kim, Kyu-Sick;Lee, Cheul-Ho;Kim, Zin-Suh
    • Journal of Korean Society of Forest Science
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    • v.96 no.6
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    • pp.667-675
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    • 2007
  • The genetic diversity and the spatial structure in two populations of Abelia tyaihyoni in Yeongwol region were studied by employing I-SSR markers. In spite of the limited distribution and small population sizes of Abelia tyaihyoni, the amount of genetic diversity estimated at the individual level was comparable to other shrub species (S.I.=0.336, h=0.217). Genetic diversity at the genet level was very similar to that at individual level. (S.l.=0.339, h=0.219). About 18.7 percent of total variation was allocated between two populations, which was slightly higher or similar level as compared with other shrub species. Genotypic diversity estimated by the ratio of the number of genets ($N_G$) over the total number of individuals (N) and a modified Simpson's index ($D_G$) were also higher than those of other shrubs. The maximum diameter of a genet did not exceed 5.5 m. The high level of gene and genotypic diversity, and the relatively limited maximum diameter of a genet suggested that the clonal propagation is not the most dominant factor in determining the population structure of Abelia tyaihyoni. Spatial autocorrelation analysis revealed significant spatial genetic structure within 12 m and 18 m distances in two populations A and B, respectively. Autocorrelations among individuals at the both individual and genet levels in each population didn't show any considerable differences. As a sampling strategy for ex-situ conservation of populations showing continuous distribution, a minimum distance of 18 m between individuals was recommended. For the populations with many segments, it was considered very crucial to sample materials from as many segments as possible.

Assessment of genetic diversity and population structure of commercial button mushroom (Agaricus bisporus) strains in Korea (한국의 상업적 양송이 균주의 유전적 다양성 및 집단 구조)

  • Lee, Hwa-Yong;An, Hye-jin;Oh, Youn-Lee;Jang, Kab-Yeul;Kong, Won-Sik;Ryu, Ho-jin;Chung, Jong-Wook
    • Journal of Mushroom
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    • v.17 no.4
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    • pp.171-178
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    • 2019
  • Agaricus bisporus is a functional food and among the most widely cultivated mushrooms in the world. In this study, we analyzed the genetic diversity and population structure of 23 Korean and 42 foreign A. bisporus cultivars using SSR (Simple sequence repeat) markers. Genetic diversity of A. bisporus cultivars was as follows: number of alleles was approximately 13; observed and expected heterozygosity were approximately 0.59 and 0.74, respectively; and polymorphic information content value was about 0.71. A. bisporus cultivars were divided into three groups using distance-based analysis. Genetic diversity of Group 2, which consisted of cultivars from various countries, was high. In addition, model-based subpopulations were divided into two, and the genetic diversity of Pop2 (Population 2), which had many cultivars, was high. The results of this study could be used in a breeding program for A. bisporus, such as the development of new genetic resources and securing diversity.