• Title/Summary/Keyword: AFLP

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Fine-scale Spatial Genetic Structure of a Small Natural Stand of Populus davidiana in South Korea using AFLP markers (AFLP 마커를 이용한 소규모 사시나무림의 공간적 유전구조 구명)

  • Lee, Min Woo;Hong, Kyung Nak;Park, Yu Jin;Lee, Jei Wan;Lim, Hyo In
    • Journal of Korean Society of Forest Science
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    • v.105 no.3
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    • pp.309-314
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    • 2016
  • A locally adapted plant population under harsh environmental changes might survive for a long generation through maintaining proper level of genetic diversity. When it happens losing the genetic diversity too much fast, the population could be declining and probably become extinct. An isolated small population of Populus davidiana was investigated to study out the genetic diversity and the fine-scale spatial genetic structure. The estimated number of adult trees in the population of Mt. Worak, South Korea, was 350 in the total area of $14,000m^2$. The number of adults in a study plot ($70m{\times}70m$) was 123. The average age was 16-year-old and a 32-year-old tree was the oldest. The distribution of individuals was slightly aggregated in the plot. Sixty-one among the 123 individuals were randomly sampled to estimate genetic variation using AFLP markers. One hundred fifty-one (77%) of total 196 amplicons were polymorphic from six AFLP primer combinations. The average number of loci per primer combination was 32.7 (S.D.=7.2). Expected heterozygosity ($H_e$) and Shannon's diversity index (S.I.) were 0.154 and 0.254, respectively. These values were extremely lower than those of other P. davidiana populations in South Korea. Genetic patchiness was showed within 21 meters by spatial autocorrelation analysis and the isolated small size of population might be mainly attributed to the formation of such small patch size.

Amplified fragment length polymorphism fingerprinting analysis of Staphylococcus aureus isolated from bovine mastitis milk (소 유방염 유래 Staphylococcus aureus의 AFLP 지문분석)

  • Kim, Yeon-soo;Kim, Sang-kyun;Hwang, Eui-kyung
    • Korean Journal of Veterinary Research
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    • v.41 no.2
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    • pp.157-165
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    • 2001
  • Amplified fragment length polymorphism(AFLP) technique is based on the polymorphism detection through selective PCR amplification of restriction fragments from digested genomic DNA and thus includes the procedures of the total DNA digestion by endonucleases, ligation of adapters to the ends of the fragments, and following the selective amplification of the restricted DNA fragments. This study were aimed to : (1) determine the genetic variability of S aureus strains, (2) estimate genetic diversity within and among these strains, (3) compare phylogenetic relationships among these strains as genetic markers using AFLP techniques. Genomic DNA was digested with a particular combination of three restriction enzymes with specific recognition sites and the DNA fragments were ligated to restriction specific adapters and amplified using the selective primer combinations. In the S aureus strain, the number of scorable AFLP bands detected per each primer combination varied from 29 to 102, with an average of 61.59 using 27 primer combinations. A total of 1,663 markers were generated, 904 bands of which were polymorphic, showing a 33.48% level of polymorphism with these primer combinations. Among the primer combinations, E02/T02, E02/T03, E04/H02, E02/T01 and E04/H03 primer combinations showed a high level of polymorphism with 0.78, 0.76, 0.74, 0.71 and 0.70, respectively. But T03/H01, E01/T02 and E01/T03 primer combinations showed a low level of polymorphism with 0.38, 0.37 and 0.15, respectively, Therefore, the former primer combinations will be the most effective for AFLP analysis of S aureus. In SA1 sub-types the level of polymorphism of S aureus KCTC 1927 was similar to that of S aureus CU 01(0.825) and higher than those of other strains such as S aureus CU 02 (0.715), S aureus KCTC 2199(0.625), S aureus KCTC 1916(0.607) and S aureus KCTC 1621 (0.553). In SA2 sub-types the level of polymorphism of S aureus CU 07 was similar to that of S aureus CU 08(0.935) and higher than those of both S aureus CU 04(0.883) and S aureus CU 05(0.883) and lower than those of S aureus CU 03(0.583). In SA3 subtypes the level of polymorphism of S aureus CU 11 was similar to that of S aureus CU 12(0.913) and lower than that of S aureus CU 15(0.623). The results proved that AFLP marker analysis of S aureus strain could be used to study the epidemiology of mastitis and in addition, common genotype in geographic region could be useful for the development of an effective vaccine or DNA marker for easy diagnosis of mastitis caused by S aureus infection.

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Morphological Variations and Genetic Variations Inferred from AFLP (Amplified Fragment Length Polymorphism) Analysis of Cottus Populations (Scorpaeniformes: Cottidae) in Korea (한국산 둑중개속 어류 개체군들의 형태 변이 및 AFLP 분석을 통한 유전 변이)

  • Byeon, Hwa Kun;Kim, Keun-Sik;Song, Ha-Yoon;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.21 no.2
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    • pp.67-75
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    • 2009
  • Morphological and genetic variation of Cottus populations in Korea were compared to each other in the watershed. Morphological variation was analyzed by meristic and morphometric characters, and by fertilized egg sizes. Genetic variation was assessed by amplified fragment length polymorphism (AFLP) fingerprinting. Cottus koreanus populations of rivers and streams running towards the eastern coasts of the Korean Peninsula were significantly different in morphometric characters from populations in rivers and streams running towards the western and southern coasts of the Korean Peninsula, but did not differ in meristric characters and fertilized egg sizes. Cottus populations in the upper streams-Yangyangnamadae Stream, Jusu Stream and Samcheogosip Stream-were the same in meristic and genetic characters. However, the Cottus sp. population from Baebong Stream is related to C. hangiongensis in meristic characteristics and to C. koreanus in the morphometric characteristics of length of the ventral fin ray and in the fertilized egg size. Pairwise genetic distances assessed by the AFLP method among C. koreanus populations were in the range of 0.110 to 0.221. Genetic distances between C. hangiongensis and C. koreanus populations varied from 0.542 to 0.621, and those between the Cottus sp. population of Baebong Stream and C. koreanus populations from 0.222 to 0.304. The result of the UPGMA dendrogram shows the Cottus sp. population of Baebong Stream was clearly separated of other C. koreanus populations.

Genetic Diversity and Population Structure of the Endangered Fish Pseudopungtungia nigra (Cyprinidae) from the Geum and Mankyung Rivers Assessed by Amplified Fragment Length Polymorphism (금강과 만경강에 서식하는 멸종위기 어류 감돌고기 Pseudopungtungia nigra의 AFLP에 의한 유전 다양성 및 집단구조)

  • Kim, Keun-Sik;Yun, Young-Eun;Kang, Eon-Jong;Yang, Sang-Geun;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.21 no.2
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    • pp.76-80
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    • 2009
  • Genetic diversity and genetic structure within the Geum River and Mankyung River populations of the Korean endangered Black shinner (Pseudopungtungia nigra) were assessed by amplified fragment length polymorphism (AFLP). AFLP analysis using five primer combinations generated 447 AFLP bands with 64.1% polymorphism (Geum River 74.6% and Mankyung River 53.6%). The heterozygosities within the two populations were calculated to be 0.170 and 0.104, respectively. Their average genetic diversities are 0.240 and 0.147, respectively. The pairwise Fst value (0.150) indicated distinct genetic differentiation between the two populations. A UPGMA dendrogram based on genetic distance among the individuals revealed a division corresponding to geographical regions, with low genetic variation within the Mankyung River population, and low genetic distance (0.026) between the two populations. Consequently, the two populations may have the same genetic origin The Geum River population will be more suitable than the Mankyung River population for conservation plans to increase the population sizes. Genetic and habitat management will be necessary for the Mankyung River population.

Genetic Diversity of an Endangered Fish, Iksookimia choii (Cypriniformes), from Korea as Assessed by Amplified Fragment Length Polymorphism (AFLP 분석에 의한 멸종위기어류 미호종개, Iksookimia choii의 유전 다양성)

  • Lee, Il-Ro;Lee, Yoon-A;Shin, Hyun-Chur;Nam, Yoon-Kwon;Kim, Woo-Jin;Bang, In-Chul
    • Korean Journal of Ecology and Environment
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    • v.41 no.1
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    • pp.98-103
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    • 2008
  • Genetic diversity and population genetic structure within or among three stream populations (Gab, Baekgok and Ji streams) of Korean endangered natural monument fish, Iksookimia choii, were assessed by amplified fragment length polymorphism (AFLP). AFLP analysis using three primer combinations generated 104 to 106 AFLP bands, and percent polymorphic bands were similar in those three populations ranging 21.5 to 24.5%. Heterozygosity and genetic diversity within or among populations were quite low for all of these populations with average values ranging from 0.067 to 0.084 and from 0.076 to 0.087, respectively. Analyses of pairwise distance and genetic similarity among three populations of I. choii also revealed the similar results with very low genetic differentiation one another. Although pairwise Fst values were very low, our data clearly indicated distinct genetic differentiation among the three populations. This is the first report concerning the genetic diversity and differentiation of this species, and provides basic genetic information that should facilitate attempts to conserve this species.

Analysis of Genetic Relationship of Pear (Pyrus spp.) Germplasms Using AFLP Markers (AFLP 표지를 이용한 배 유전자원의 유연관계 분석)

  • Cho, Kang-Hee;Shin, Il Sheob;Kim, Hyun Ran;Kim, Jeong-Hee;Heo, Seong;Yoo, Ki Yeol
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.444-450
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    • 2009
  • Amplified fragment length polymorphism (AFLP) marker was utilized for evaluation of genetic diversity of 60 pear germplasms. Twenty selective AFLP primer pairs generated a total of 522 polymorphic amplification products. From UPGMA (unweighted pair-group method arithmetic average) cluster analysis by using polymorphic bands, the pear germplasms were divided into four clusters by similarity index of 0.691. The first cluster (I) included European pears belonging to Pyrus communis and wild species such as P. nivalis and P. cordata. The second cluster (II) included Ussurian pea pears belonging to P. betulaefolia and P. fauriei. The third cluster (III) included pea pears belonging to P. calleryana and P. koehnei. Most of germplasms belonging to P. pyrifolia and P. ussuriensis, and interspecific hybrids were included in the fourth (IV) cluster. Therefore pear germplasms originated from East Asia were closely related to P. pyrifolia and P. ussuriensis. Similarity values among the tested pear germplasms ranged from 0.584 to 0.879, and the average similarity value was 0.686.

Application of AFLPs to Phylogenetic Analysis of Aegilops (AFLPs에 의한 Aegilops의 계통발생학적 재평가)

  • Park, Yong-Jin;Shim, Jae-Wook
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.6
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    • pp.790-799
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    • 1997
  • Aegilops genus is known to include the donor species of the Band D genome of the bread wheat(ABD). An effort to establish a better strategy for phylogenetic relationships about Aegilops polyploids by AFLPs(Amplified Fragment Length Polymorphisms) was conducted using the 19 Aegilops sPP. and T. aestivum. The 207 polymorphic bands from the amplified products on the 6% acrylamide denaturing sequencing gels were obtained with the 7 AFLP primer combinations, and used to account for the genetic similarities and cluster analysis using NTSYS program. According to the genome analysis, the $M^h$-genome of Ae. heldreichii was estimated as an intermediate genome between the M-genome of Ae. comosa and N-genome of Ae. uniaristata and supposed to be incorporated in the establishing process of UM-genome as a possible diploid donor. And Ae. ventricosa(DN) was more close to Ae. umbellulata(U) than Ae. squarrosa(D). The close relationship between Ae. squarrosa and T. aestivum was perceived as a diploid donor of D-genome. As for the polyploid species, hexaploid Ae. triaristata was more closely related to Ae. columnaris rather than tetraploid Ae. triaristata. The clustered groups were, basically same to the previous Gihara's sections based on phenotypes and pairing analysis of interspecific hybrids. AFLP was evaluated as an efficient and powerful method in the genome evaluation of closely related species.

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Application of Chromosome Manipulation, DOP-PCR and AFLP Methods to Isolate Sex-Specific DNAs from Rumex acetosa L.

  • Jin, Dong-Chung;Kim, Joong-Soon;Park, ji-Young;Bong, Jae-Wook;Hur, Yoon-Kang
    • Journal of Photoscience
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    • v.12 no.2
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    • pp.75-82
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    • 2005
  • Rumex acetosa L. is a dioecious flowering plant with well developed sex chromosome system: 2n = 12 + XX in the female plants and 2n = 12 + XY1Y2 in the male plants. To isolate sex-linked DNA, we carried out chromosome micromanipulation, followed by DOP-PCR, AFLP of the PCR products, reverse Southern hybridization and sequence analysis. From 500 AFLP specific clones, 13 X-chromosome and 5 Y-chromosome specific clones were obtained. Except one clone RADAX-239 ($\underline{R}umex\;\underline{a}-\underline{D}OP-PCR-\underline{A}FLP-\underline{Y}-chromosome\;specific$), all clones appear to be R. acetosa plant-specific sequences and non-coding sequences. Southern blot analysis using these clones could not discriminate genomic DNAs either from male or female plants. Results of this study imply that both autosome-origin and degeneration of sex chromosomes are prevalent in plant systems.

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Determination of the minimum population size for ex situ conservation of water-shield (Brasenia schreberi J.F. Gmelin) inferred from AFLP analysis

  • Kim, Changkyun;Na, Hye Ryun;Jung, Jongduk;Kim, Hojoon;Hyun, Jin-Oh;Shin, Hyunchur;Choi, Hong-Keun
    • Journal of Ecology and Environment
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    • v.35 no.4
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    • pp.301-306
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    • 2012
  • Determination of the minimum population size is an important component for the ex situ conservation of an endangered species. Here, we present the identification of collection strategies that most efficiently captured the genetic diversity of Brasenia schreberi J.F. Gmelin (water-shield) in natural populations from the mainland (MGC) and Jeju Island (JNS) of South Korea, using amplified fragment length polymorphism (AFLP) markers. A total of 313 and 383 polymorphic bands were detected in the MGC and JNS populations, respectively. All of the 140 sampled ramets were distinguishable by the presence of distinct AFLP phenotypes. According to the simulation of the individual sampling by maximization sampling, 25 and 28 individuals captured all of the genetic diversity in the MGC population (mainland of South Korea) and the JNS population (Jeju Island), respectively. The level of genetic diversity of the core collections was similar to the entire collection, indicating that the core collections very well represent the diversity of the entire collection. We therefore suggest a management unit of B. schreberi based on the genetic information for assessing the minimum population size for its ex situ conservation.

A Fluorescence-based cDNA-AFLP Method for Identification of Differentially Expressed Genes

  • Park, Sook-Young;Jwa, Nam-Soo;Chi, Myoung-Hwan;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.25 no.2
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    • pp.184-188
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    • 2009
  • Identification of differently expressed genes under specific tissues and/or environments provides insights into the nature and underlying mechanisms of cellular processes. Although cDNA-AFLP (Amplified Fragment Length Polymorphism) is a powerful method for analyzing differentially expressed genes, its use has been limited to the requirement of radioactive isotope use and the difficulty of isolating the bands of interest from a gel. Here, we describe a modified method for cDNA-AFLP that uses a fluorescence dye for detection and isolation of bands directly from a small size polyacrylamide gel. This method involves three steps: (i) preparation of cDNA templates, (ii) PCR amplification and differential display, and (iii) identification of differentially expressed genes. To demonstrate its utility and efficiency, differentially expressed genes during vegetative growth and appressorial development of Magnaporthe oryzae were analyzed. This method could be applied to compare gene expression profiles in a diverse array of organisms.