• Title/Summary/Keyword: AFLP analysis

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Diversity and Inheritance of AFLP Markers in Wild and Cultivated Soybeans (AFLP marker를 이용한 콩의 유전적 다양성과 유전분리 분석)

  • 김용호;윤홍태
    • Korean Journal of Plant Resources
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    • v.17 no.3
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    • pp.265-271
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    • 2004
  • Genetic variation is the basis of crop improvement. Limited genetic diversity in a crop species may restrict the amount of genetic improvement that can be achieved through plant breeding. Soybean is one of the world's most important crops. A potential source of genetic variability for the cultivated soybean is the wild species G. soja Sieb. & Zucc. Amplified fragment length polymorphism (AFLP) analysis is a PCR-based technique, which can detect a 10-fold greater nubmer of loci than other DNA marker analysis. Twenty cultivated soybeans and two-hundred wild soybeans were used to determine genetic vatiations by AFLPs and evaluate the usefulness of AFLPs as DNA markers. Six-hundred and ten fragments were detected with an average of 56 AFLP fragments produced per primer in a total of 11 AFLP primer pairs. The number of polymorphic loci detected per primer ranged from 7 to 20 and the polymorphism was greater in wild than in cultivated soybean. F$_2$ segregation analysis of four AFLP fragments in combination of Hwaeomputkong ${\times}$ PI 417479 indicated that they segregate as stable Mendelian loci with 3 : 1. This results strongly suggest that the AFLP analysis is a good technique for the detection of genetic polymorphism in a wide plant species.

Amplified Fragment Length Polymorphism Fingerprinting as a Tool to Study the Genetic Diversity of Staphylococcus aureus Isolated from Food Sources

  • Kim, Young-Sam;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.8 no.1
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    • pp.39-46
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    • 2002
  • Amplified fragment length polymorphism (AFLP) is a recently developed PCR-based high resolution fingerprinting method that is able to generate complex banding patterns which can be used to delineate intraspecific genetic relationships among bacteria. In this study, we have modified and evaluated a PCR-based technique, amplified fragment length polymorphism (AFLP) analysis, for use in fingerprinting strains of Staphylococcus aureus. Single-enzyme amplified fragment length polymorphism (SE-AFLP) analysis was used to perform strain identification of Staphylococus aureus. By careful selection of AFLP primers, it was possible to obtain reproducible and sensitive identification to strain level. AFLP fingerprinting of 5 reference strains of Staphylococcus aureus and 65 strains of Staphylococcus aureus that were isolated from food sources of different area and diverse genomic types of Staphylococcus aureus were recognized. As a result of this study, we found that the AFLP patterns of Staphylococcus aureus isolated from Seoul, Taejeon and Gwang-Ju indicated the close relation with genetic similarity. The main purpose of this study was to find an alternative and reliable fingerprinting method to study the overall genetic diversity, using Staphylococcus aureus species as an example, and observed if the method can be successfully applied to all staphylococcal species.

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Fingerprinting of Listeria monocytogenes by Amplified Fragment Length Polymorphism Analysis

  • Jin, Hyun-Seok;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.8 no.1
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    • pp.29-37
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    • 2002
  • Listeria monocytogenes poses an increasing health risk, which in part is due to increasing health risk, consumption of ready-to-eat food products and the introduction of increasing numbers of food products from regions with different dietary habits. L. monocytogenes can be present in meat, shellfish, vegetables, unpasteurised milk and soft cheese and poses a risk if food containing these products is stored at refrigeration temperature and is not properly heated before consumption, as L. monocytogenes is psychrophilic. Amplified-fragment length polymorphism (AFLP) analysis is the method of genotypic techinique in which adaptor oligonucleotides are ligated to restriction enzyme fragments and then used as target sites for primers in a PCR amplification. The amplified fragments are electrophoretically separated to give strain-specific band profiles. Single-enzyme approach that did not require costly equipment or reagents for the fingerprinting of strains of Listeria monocytogenes was developed. Single-enzyme amplified fragment length polymorphism (SE-AFLP) analysis was used to perform species and strain identification of Salmonella, Shigella, Yersinia and E. coli. By careful selection of AFLP primers, it was possible to obtain reproducible and sensitive identification to strain level. The AFLP patterns of L. monocytogenes are divided by the kinds of specimens in which were isolated. SE-AFLP fragments can be analyzed using standard gel electrophoresis, and can be easily scored by visual inspection, due to the low complexity of the fingerprint obtained by this method. These features make SE-AFLP suitable for use in either field or laboratory applications.

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Genetic Diversity of Curcuma Genus Collected Germplasm using Analysis of AFLP (AFLP 분석을 통한 Curcuma속 수집종간 유전적 다양성 분석)

  • Moon, Byeong Cheol;Kim, Wook Jin;Ji, Yunui;Lee, Young Mi;Kim, Ho Kyoung
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.6
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    • pp.455-460
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    • 2013
  • Collected germplasms of five representative species belonging to Curcuma genus (C. longa, C. aromatica, C. zedoaria, C. phaeocaulis and C. kwangsiensis) were 52 samples from different farmhouse in Korea and China. To elucidate the genetic diversity among the species, 52 samples were analyzed by genomic fingerprinting method using amplified fragment length polymorphism (AFLP). AFLP results of 6 primer combinations were revealed 643 total DNA fragments and 349 polymorphic bands with the 54.3% ratio of polymorphism. In the analysis of coefficient similarity using unweight pair group method with arithmetic averages (UPGMA), 52 Curcuma germplasm lines were ranged from 0.60 to 0.99 and clustered distinct five groups according to the species and collected geographical levels. However, the result of principal coordinate analysis (PCA) by multi-variate analysis was shown significantly greater differences among species than geographical origins based on AFLP profiling data of these samples.

Comparative AFLP Profiles among Strains of Korean Races of Xanthomonas oryzae pv. oryzae.

  • Kang, Mi-Hyung;Lee, Du-Ku;Noh, Tae-Hwan;Shim, Hyeong-Kwon;Na, Seung-Yong;Kim, Jae-Duk
    • Plant Resources
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    • v.7 no.1
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    • pp.65-68
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    • 2004
  • We used an amplified fragment length polymorphism (AFLP) analysis, a novel PCR-based technique, to differentiate Xanthomonas oryzae pv. oryzae (Xoo) of Korean races. The 6 strains of Xoo K1, K2, K3 races were tested with 81 AFLP primer combinations to identify the best selective primers. The primer combinations were selected according to their reproducibility, number of polymorphic bands and polymorphism detected among Xoo strains. 18 strains of Xoo K1, K2 and K3 races were analyzed with the selected combinations of primer set. Some primer combinations (Eco R I +1 / Mse I+1) could differentiate Xoo of Korean races that were not distinguished by other fingerprinting analysis. Thus AFLP fingerprinting permitted very fine discrimination among different races.

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Genetic Variation of Rhododendron micranthum Based on AFLP and RAPD Analysis (AFLP와 RAPD 방법을 이용한 꼬리진달래(Rhododendron micranthum) 수집종의 유전적 변이 분석)

  • 김남수;김진홍;이주경;김남희;이명숙;이재선;박철호
    • Korean Journal of Plant Resources
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    • v.17 no.3
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    • pp.227-238
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    • 2004
  • Rhododendron micranthum is an endangered species in Korea. In order to develop the strategy of gene diversity conservation, estimation of the amount of genetic diversity, the genetic variation and relationship in the native populations of Rh. micranthum was performed on the basis of AFLP and RAPD analysis. Analysis of 56 accessions derived from 6 populations of Rh. micranthum with four AFLP primer combinations and ten RAPD primers detected a total of 33 polymorphic AFLP fragments and 15 polymorphic RAPD fragments, respectively. By UPGMA cluster analysis with molecular markers, the 56 accessions were grouped into three major clusters at 73.3% genetic similarity; group I consists of most accessions of populations I, II, IV, V and Ⅵ, group II consists of 7 accessions of population III, and group III consists of only two accessions of population IV. The geographic locations of the most accessions derived from six populations were not related to their position in the UPGMA cluster analysis, except for several accessions of populations III and IV. The genetic similarity of among six populations measured by AFLP and RAPD markers ranged from 0.66 to 0.99. Among them, population Ⅵ showed the highest GS with means of 0.87, while population I showed the lowest GS with means of 0.78. This result will be useful for designing the strategy of conservation in the native populations of Rh. micranthum.

DNA Fingerprinting of Rice Cultivars using AFLP and RAPD Markers

  • Cho, Young-Chan;Shin, Young-Seop;Ahn, Sang-Nag;Gleen B. Gregorio;Kang, Kyong-Ho;Darshan Brar;Moon, Huhn-Pal
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.1
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    • pp.26-31
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    • 1999
  • This experiment was conducted to evaluate genetic variation in 48 rice accessions (Oryza sativa L.) using AFLP and RAPD markers. For AFLP, a total of 928 bands were generated with 11 primer combinations and 327 bands (35.2%) of them were polymorphic among 48 accessions. In RAPD analyses using 22 random primers 145 bands were produced, and 121 (83.4%) were polymorphic among 48 accessions. Each accession revealed a distinct fingerprint by two DNA marker systems. Cluster analysis using AFLP-based genetic similarity tended to classify rice cultivars into different groups corresponding to their varietal types and breeding pedigrees, but not using RAPD-based genetic similarity. The AFLP marker system was more sensitive than RAPD in fingerprinting of rice cultivars with narrow genetic diversity.

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Cloning and Characterization of Genes Controlling Flower Color in Pharbitis nil Using AFLP (Amplified Fragment Length Polymorphism) and DDRT (Differential Display Reverse Transcription)

  • Kim, Eun-Mi;Jueson Maeng;Lim, Yong-Pyo;Yoonkang Hur
    • Journal of Photoscience
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    • v.7 no.2
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    • pp.73-78
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    • 2000
  • To analyze molecular traits determining pigmentation between Pharbitis nill violet and white, Amplified Fragment Length Polymorphism(AFLP) and Differential Display Reverse Transcription(DDRT) experiments were carried out with either genomic DNAs or total RNAs isolated from both plants. Results of AFLP experiment in combination of 8 EcoRⅠ primers with 6 MseⅠ primers showed 41 violet-and 60 white-specific DNA bands. In the subsequent experiment, 22 violet-and 22 white-specific DNA fragments were amplified by PCR with DNAs eluted. The sizes of the fragments range from 200 to 600bp. DDRT using total RNA produced 19 violet-and 17 white-specific cDNA fragments, ranging from 200 to 600bp. The fragments obtained by both AFLP and DDRT had been cloned into pGEM T-easy vector, amplified and subjected to the nucleotide sequence analyses. As a result of Blast sequence analysis, most of them sequenced up to date showed no similarity to any Known gene, while few has similarity to known animal or plant genes. An AFLP clone V6, for example, has a strong sequence similarity to the human transcription factor LZIP-alpha mRNA and a DDRT clone W19 to Solanum tuberosum 3-hydroxy-3-methylglutaryl coenzyme A reductase mRNA.

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Genetic Relationships among Korean Adlay, Coix lachryma-jobi L., Landraces Based on AFLPs

  • Moon Jung-Hun;Jang Jung Hee;Park Jung Soo;Kim Sung Kee;Lee Kyung-Jun;Lee Sang-Kyu;Kim Kyung-Hee;Lee Byung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.2
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    • pp.142-146
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    • 2005
  • Thirty-two germplasms of Korean adlay landraces were examined to analyse the genetic relationship through the amplified fragment length polymorphism (AFLP) approach. Total number of AFLP products generated by 12 selective primer combinations was 882. The number of polymorphic fragments by each primer combination greatly varied from 4 to 51 with a mean of 20.3, bands visible on the polyacrylamide gel. A genetic similarity coefficient was used for cluster analysis following UPGMA (unweighted pair grouping method of averages) method. The resulting clusters were represented in the form of a dendrogram. The clustering was not tight in the dendrogram. There was generally no clear grouping of the adlay according to the geographic regions in which germplasms were collected. The present AFLP analysis imply that although Korean adlay displayed a larger amount of AFLP variation within germplasms, the variation was shown independently without reflecting a clinal variation. This study demonstrated that AFLP method can be used to examine the genetic relationships among different germplasms of adlay.

Genetic Variation of Monilinia fructicola Population in Korea

  • Su In Lee;Hwa-Jung Lee;Youn-Sig Kwak
    • The Plant Pathology Journal
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    • v.40 no.2
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    • pp.205-217
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    • 2024
  • Brown rot disease, caused by Monilinia spp., poses a significant threat to pome and stone fruit crops globally, resulting in substantial economic losses during pre- and post-harvest stages. Monilinia fructigena, M. laxa, and M. fructicola are identified as the key agents responsible for brown rot disease. In this study, we employed the amplified fragment length polymorphism (AFLP) method to assess the genetic diversity of 86 strains of Monilinia spp. isolated from major stone fruit cultivation regions in South Korea. Specifically, strains were collected from Chungcheong, Gangwon, Gyeonggi, Gyeongsang, and Jeolla provinces (-do). A comparative analysis of strain characteristics, such as isolation locations, host plants, and responses to chemical fungicides, was conducted. AFLP phylogenetic classification using 20 primer pairs revealed the presence of three distinct groups, with strains from Jeolla province consistently forming a separate group at a high frequency. Furthermore, M. fructicola was divided into three groups by the AFLP pattern. Principal coordinate analysis and PERMANOVA were applied to compare strain information, such as origin, host, and fungicide sensitivity, revealing significant partition patterns for AFLP according to geographic origin and host plants. This study represents the utilization of AFLP methodology to investigate the genetic variability among M. fructicola isolates, highlighting the importance of continuous monitoring and management of variations in the brown rot pathogen.