• Title/Summary/Keyword: Simple sequence repeats

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Genetic Structure of Macrophomina phaseolina Populations, the Causal Agent of Sesame Charcoal Rot Disease in Iran

  • Maryam Dolatkhah;Fariba ghaderi;Abdollah Ahmadpour
    • Research in Plant Disease
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    • v.30 no.1
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    • pp.50-59
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    • 2024
  • Charcoal rot disease, caused by the fungus Macrophomina phaseolina, is one of the most important diseases of Sesame (Sesamum indicum) all over the world. However, the population biology of M. phaseolina is poorly understood. In this study, M. phaseolina isolates from five different regions of Iran (Khuzestan, Fars, Bushehr, Hormozgan, and Kohgiluyeh & Boyer-Ahmad provinces) (n=200) were analyzed for genetic variation using inter simple sequence repeats marker. In total, 152 unique haplotypes were identified among the 200 M. phaseolina isolates, and gene diversity (H=0.46-0.84) and genotypic diversity were high in each of the regions. The structure analysis clustered five Iranian populations into two distinct groups, the individuals from group 1 were assigned to the Bushehr population and the individuals from Khuzestan, Fars, Hormozgan and Kohgiluyeh & Boyer-Ahmad were aggregated and formed group 2. The results matched with genetic differentiation and gene flow among regions. Analyses of the distribution of gene diversity within and among five Iranian populations were 61% and 39%, respectively. Our results showed that infected seeds are thought to be the dominant mechanism responsible for the spreading of the pathogen in southern parts of Iran. In summary, it is essential to have local quarantine and prevent seed exchanges between geographical populations to restrict the dispersal of pathogen over long distances and provide certified seeds in Iran.

Applications of molecular markers in the discrimination of Panax species and Korean ginseng cultivars (Panax ginseng)

  • Jo, Ick Hyun;Kim, Young Chang;Kim, Dong Hwi;Kim, Kee Hong;Hyun, Tae Kyung;Ryu, Hojin;Bang, Kyong Hwan
    • Journal of Ginseng Research
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    • v.41 no.4
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    • pp.444-449
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    • 2017
  • The development of molecular markers is one of the most useful methods for molecular breeding and marker-based molecular associated selections. Even though there is less information on the reference genome, molecular markers are indispensable tools for determination of genetic variation and identification of species with high levels of accuracy and reproducibility. The demand for molecular approaches for marker-based breeding and genetic discriminations in Panax species has greatly increased in recent times and has been successfully applied for various purposes. However, owing to the existence of diverse molecular techniques and differences in their principles and applications, there should be careful consideration while selecting appropriate marker types. In this review, we outline the recent status of different molecular marker applications in ginseng research and industrial fields. In addition, we discuss the basic principles, requirements, and advantages and disadvantages of the most widely used molecular markers, including restriction fragment length polymorphism, random amplified polymorphic DNA, sequence tag sites, simple sequence repeats, and single nucleotide polymorphisms.

Phylogenic Study of Genus Phyllostachys (Phyllostachys) in Korea by Internal Transcribed Spacer Sequence (ITS) (ITS 서열에 의한 한국 왕대속 식물종의 계통분류학적 연구)

  • Lee, Song-Jin;Huh, Man-Kyu;Huh, Hong-Wook;Lee, Byeong-Ryong
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1281-1287
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    • 2011
  • Phyllostachys consists of high and fast growing trees and is a genus in the family Gramineae. The genus has many species in Asia, with main distribution being in India and China. One of the most popular sequences for phylogenetic inference at the generic and infrageneric levels in plants is the internal transcribed spacer (ITS) region of the 18S-5.8S-26S nuclear ribosomal cistron. We evaluated four taxa with the ITS region to estimate phenotypic relationships within the genus Phyllostachys in Korea. Alignment of the DNA sequences required the addition of numerous gaps. Sequence variation within the Phyllostachys was mostly due to natural selection, although several indels and inserts were found. Within the genus Phyllostachys, P. nigra and P. nigra var. henonis were the relatives in the three phylogenetic analyses (MP, ML, and NJ). However, some external nodes were poorly supported. Morphological traits and simple repeats (ISSR) represented the result of a relationship similar to the that of ITS sequences in the genus Phyllostachys. This suggests that ITS sequences are very informative for identification of these taxa.

Development of Multiplex Microsatellite Marker Set for Identification of Korean Potato Cultivars (국내 감자 품종 판별을 위한 다중 초위성체 마커 세트 개발)

  • Cho, Kwang-Soo;Won, Hong-Sik;Jeong, Hee-Jin;Cho, Ji-Hong;Park, Young-Eun;Hong, Su-Young
    • Horticultural Science & Technology
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    • v.29 no.4
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    • pp.366-373
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    • 2011
  • To analyze the genetic relationships among Korean potato cultivars and to develop cultivar identification method using DNA markers, we carried out genotyping using simple sequence repeats (SSR) analysis and developed multiplex-SSR set. Initially, we designed 92 SSR primer combinations reported previously and applied them to twenty four Korean potato cultivars. Among the 92 SSR markers, we selected 14 SSR markers based on polymorphism information contents (PIC) values. PIC values of the selected 14 markers ranged from 0.48 to 0.89 with an average of 0.76. PIC value of PSSR-29 was the lowest with 0.48 and PSSR-191 was the highest with 0.89. UPGMA clustering analysis based on genetic distances using 14 SSR markers classified 21 potato cultivars into 2 clusters. Cluster I and II included 16 and 5 cultivars, respectively. And 3 cultivars were not classified into major cluster group I and II. These 14 SSR markers generated a total of 121 alleles and the average number of alleles per SSR marker was 10.8 with a range from 3 to 34. Among the selected markers, we combined three SSR markers, PSSR-17, PSSR-24 and PSSR-24, as a multiplex-SSR set. This multiplex-SSR set used in the study can distinguish all the cultivars with one time PCR and PAGE (Polyacrylamide gel electrophoresis) analysis and PIC value of multiplex-SSR set was 0.95.

Inter Simple Sequence Repeats (ISSR) Marker Analysis of Genetic Diversity in Korean Phasianus colchicus karpowi and Genetic Relationships Among Subspecies of Phasianus spp. (Inter Simple Sequence Repeats (ISSR) 표지자를 이용한 한국꿩의 유전적 다양성 및 아종간의 유연관계 분석)

  • Yoon, Seong-Il
    • Korean Journal of Environmental Biology
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    • v.26 no.2
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    • pp.66-75
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    • 2008
  • The level of genetic diversity and genetic relationships among Korean ring-necked pheasant (Phasianus colchicus karpowi) habitat and subspecies have been investigated based on Inter Simple Sequence Repeat (ISSR) markers. Wild and domesticated Korean ring-necked pheasant, hybrids between domesticated Korean ring-necked and foreign subspecies, and four foreign subspecies; Chinese ring-necked (P. c. torquatus), Melanistic mutant (P. c. mut. tenebrosus), XL White (P. c. mut) and Southern green (P. c. versicolor) were used for comparison. On the basis of the results of AMOV A, 94.08% of genetic diversity in Korean ring-necked was allocated among individuals within habitat differences. Estimate of $\Phi$st, which represents the degree of genetic differentiation among habitats was 5.9%. Based on the dendrogram reconstructed by UPGMA, Yangpyung habitat of the eight habitats turned out to be distinct from others habitat. Interestingly, domesticated Korean ring-necked and hybrid mixture showed closer genetic relationship with four foreign subspecies than Korean ring-necked. As a consequence of AMOVA, 96.63% of genetic diversity in four foreign subspecies was allocated among individuals within subspecies. Estimate of $\Phi$st representing the degree of genetic differentiation among subspecies was 3.4%, which was lower than that among habitats of Korean ring-necked. The lower level of genetic difference among four foreign subspecies showed that these subspecies were genetically closer even though they were morphologically classified into four different subspecies. When seven habitats of Korean ring-necked pheasant and four foreign subspecies were divided into Korean and Foreign Pheasant Groups, respectively, more than 17% of genetic diversity was allocated between groups (about 4% among habitats/subspecies within groups). This observation implied that Korean ring-necked pheasant is genetically quite different from four foreign subspecies. On the basis of cluster analysis, three foreign subspecies (Chinese ring-necked pheasant, Melanistic mutant pheasant, and XL White pheasant) formed a distinct group with domesticated Korean ring-necked pheasant and hybrid mixture at 98% confidence interval.

Genome Survey and Microsatellite Marker Selection of Tegillarca granosa (꼬막(Tegillarca granosa)의 유전적 다양성 분석을 위한 드래프트 게놈분석과 마이크로새틀라이트 마커 발굴)

  • Kim, Jinmu;Lee, Seung Jae;Jo, Euna;Choi, Eunkyung;Kim, Hyeon Jin;Lee, Jung Sick;Park, Hyun
    • Journal of Marine Life Science
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    • v.6 no.1
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    • pp.38-46
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    • 2021
  • The blood clam, Tegillarca granosa, is economically important in marine bivalve and is used in fisheries industry among western Pacific Ocean Coasts especially in Korea, China, and Japan. The number of chromosomes in the blood clam is known as 2n=38, but the genome size and genetic information of the genome are not still clear. In order to predict the genomic size of the T. granosa, the in-silico analysis analysed the genomic size using short DNA sequence information obtained using the NGS Illumina HiSeq platform. As a result, the genomic size of T. granosa was estimated to be 770.61 Mb. Subsequently, a draft genome assembly was performed through the MaSuRCA assembler, and a simple sequence repeat (SSR) analysis was done by using the QDD pipeline. 43,944 SSRs were detected from the genome of T. granosa and 69.51% di-nucleotide, 16.68% trinucleotide, 12.96% tetra-nucleotide, 0.82% penta-nucleotide, and 0.03% hexa-nucleotide were consisted. 100 primer sets that could be used for genetic diversity studies were selected. In the future, this study will help identify the genetic diversity of T. granosa and population genetic studies, and further identify the classification of origin between homogenous groups.

Linkage Analysis of both RAPD and I-SSR Markers using Haploid Genome from a Single Tree of Pinus densiflora S. et Z. (소나무 단일(單一) 모수(母樹)의 반수체(半數體) 게놈을 이용(利用)한 RAPD 및 I-SSR 표식자(標識子)의 연관분석(連關分析))

  • Hong, Yong-Pyo;Chung, Jae-Min;Kim, Yong-Yul;Jang, Suk-Seong
    • Journal of Korean Society of Forest Science
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    • v.89 no.4
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    • pp.536-542
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    • 2000
  • A linkage map for Japanese red pine (Pinus densiflora) was constructed on the basis of two DNA marker systems of random amplified polymorphic DNAs (RAPDs) and inter-simple sequence repeats (I-SSR). Haploid genomic DNAs were extracted from megagametophyte tissues of 96 individual seeds in a single tree. A total of 98 DNA markers including 52 RAPD markers amplified by 25 primers and 46 I-SSR markers amplified by 18 primers were verified as Mendelian loci showing 1 : 1 segregation in 96 megagametophytes which were ${\chi}^2$-tested at 5% significance level. Of them, 63 segregating loci turned out to be linked into 20 linkage groups by the two-point analysis. However, 35 loci (17 RAPD and 18 I-SSR) of the 98 segregating loci did not coalesced into any linkage groups at a LOD of 3.0. The linked 63 loci were separated by an average distance of about 25.5 cM, which were spanned 1097.8 cM as a whole. The minimum and maximum map distances of the linkage groups were 4.3 cM and 54.9 cM, respectively. Incorporation of I-SSR loi into linkage map of RAPD loci resulted in extended and partially more saturated linkage blocks.

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Genetic Polymorphism of Microsatellite Markers in Panax ginseng C.A. Meyer (인삼 (Panax ginseng C.A. Meyer)의 Microsatellite 마커에 대한 유전적 다형성과 특성 규명)

  • Park, Sun-Wha;Hyun, Young-Se;Chung, Ki-Wha
    • Journal of Ginseng Research
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    • v.33 no.3
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    • pp.199-205
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    • 2009
  • Ginseng (Panax ginseng C.A. Meyer) is one of the most important medicinal plants in East Asia. Microsatellite or simple sequence repeat (SSR) markers are used in obtaining genetic analysis and authentication in many plants. The present study examined five microsatellites in conjunction with P. ginseng in Korea. The total observed allelic number was 17 (mean = 3.4), and gene diversities varied from 0.078 to 0.543 with an average of 0.314. Through a combined analysis of five loci in 100 ginseng samples, 44 different combined genotypes were observed. Expected and observed heterozygosites ranged from 0.077 to 0.541 (mean = 0.313) and 0.040 to 0.130 (0.083), respectively. All examined loci exhibited deficiency of heterozygosity and deviation from the Hardy-Weinberg equilibrium. Such results may be explained by the non-random mating and inbreeding that has occurred for several hundred years. These microsatellite markers could be used for the study of molecular genetics and the establishment of DNA marker database, as well as authentication of ginseng species and chromosomal mapping of QTL loci in P. ginseng.

Diversity of I-SSR Variants in the Populations of Torreya nucifera (비자나무 집단(集團)에서의 I-SSR 변이체(變異體)의 다양성(多樣性))

  • Hong, Yong-Pyo;Cho, Kyung-Jin;Kim, Yong-Yul;Shin, Eun-Myoung;Pyo, Sun-Kyung
    • Journal of Korean Society of Forest Science
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    • v.89 no.2
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    • pp.167-172
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    • 2000
  • Inter-simple sequence repeat (I-SSR) markers were analyzed from diploid genomes of 95 nutmeg trees (Torreya nucifera Siev. et Zucc.) in 5 populations. A total of 62 I-SSR amplicons were observed and 7 of them were monomorphic in 95 individuals. DNA fingerprint of each tree was verified by pooling the observed I-SSR amplicons. Most of the genetic diversity was allocated within population (90.65%) and all the populations revealed similar level of I-SSR amplicon diversity within population. Degree of population differentiation (${\phi}_{ST}=9.35%$) was moderate on the basis of criteria obtained from isozyme analysis. Based on the results of the cluster analysis of UPGMA, genetic relationships among 5 populations were not coincided with the pattern of geographic distribution. Non-significant confidence interval at each node also suggests that all the nutmeg populations are genetically not much differentiated.

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Distribution and Frequency of SSR Motifs in the Chrysanthemum SSR-enriched Library through 454 Pyrosequencing Technology (국화 SSR-enriched library에서 SSR 반복염기의 분포 및 빈도)

  • Moe, Kyaw Thu;Ra, Sang-Bog;Lee, Gi-An;Lee, Myung-Chul;Park, Ha-Seung;Kim, Dong-Chan;Lee, Cheol-Hwi;Choi, Hyun-Gu;Jeon, Nak-Beom;Choi, Byung-Jun;Jung, Ji-Youn;Lee, Kyu-Min;Park, Yong-Jin
    • Journal of the Korean Society of International Agriculture
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    • v.23 no.5
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    • pp.546-551
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    • 2011
  • Chrysanthemums, often called mums or chrysanths, belong to the genus Chrysanthemum, which includes about 30 species of perennial flowering plants in the family Asteraceae. We extracted DNA from Dendranthema grandiflorum ('Smileball') to construct a simple sequence repeat (SSR)-enriched library, using a modified biotin-streptavidin capture method. GS FLX (Genome Sequencer FLX System which provides the flexibility to perform the broad range of applications) sequencing (at the 1/8 run specification) resulted in 18.83 mega base pairs (Mbp) with an average read length of 280.06 bp. Sequence analyses of all SSR-containing clones revealed a predominance of di-nucleotide motifs (16,375, 61.5%) followed by tri-nucleotide motifs (6,616, 24.8%), tetra-nucleotide motifs (1,674, 6.3%), penta-nucleotide motifs (1,283, 4.8%), and hexa-nucleotide motifs (693, 2.6%). Among the di-nucleotide motifs, the AC/CA class was the most frequently identified (93.5% of all di-nucleotide types), followed by the GA/AG class (6.1%), the AT/TA class (0.4%), and the CG/GC class (0.03%). When we analyzed the distribution of different repeat motifs and their respective numbers of repeats, regardless of the motif class, of 100 SSR markers, we found a higher number of di-nucleotide motifs with 70 to 80 repeats; we also found two di-nucleotide motifs with 83 and 89 repeats, respectively, but their product lengths were within optimum size (297 and 300 bp). In future work, we will screen for polymorphisms of possible primer pairs. The results will provide a useful tool for assessing molecular diversity and investigating the population structure among and within Chrysanthemum species.