• Title/Summary/Keyword: random amplification of polymorphic DNA (RAPD)

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Genetic status of Acanthamoeba spp. Korean isolates on the basis of RAPD markers (RAPD 표지자 분석 에 의한 가시아메바속 한국분리주의 유전적 지위)

  • 홍용표;오승환
    • Parasites, Hosts and Diseases
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    • v.33 no.4
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    • pp.341-348
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    • 1995
  • Genetic status of Acnnthamoebc sap. were tested on the basis of random amplified polymorphic DNA (RAPD) marker analysis. Four previously established Accnthcmoebn species, 4 Korean isolates of Acnnthamoeba sp., and one American isolate of Acanthcmoebc sp. were analyzed by RAPD-PCR using an arbitrary decamer primers. Amplification products were fractionated by agarose gel electrophoresis and slainrd by ethidium bromide . Eighteen primers produced DNA amplification profiles revealing clear differences among 4 species. Nine of them also produced DNA amplification profiles which included some isolate-specific amplification products. On the basis of amplified fragments by 18 primers, the pairwise similarity indices between A. culbensoni and other species (i.e. A. hntchetti, A. trinngularis, A. polyphaga) were 0.300, 0.308, and 0.313, respectively. Similarity index between A. hctchetti and A. triansulcris was 0.833. The mean similarity index among the 3 Korean isolates (YM-2, -3, -4) was 0.959 and 0.832 among them and 2 other species (A. hatchetti and A. triongulnris). The mean similarity index among YM-5 and other Korean isolates (YM-2, -3, -4) was 0.237. However, the similarity index between YM-5 and A. culbeksoni was 0.857, which suggests that YM-5 is genetically more similar to A. culbertsoni than other Korean isolates. Phonogram reconstructed by UPGMA method revealed that there are two groups: one group consists of A. hctchetti, A. tlonsulcns, and 3 Korean isolates (YM-2, -3, -4) , and the other group consists of A. cuLbensoni. A. polwphosc, HOV, and YM-5.

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Genetic Relationahips of the Two Morphorogical Types of Myzus persicae(Homoptera:Aphididae) Collected from Tobacco Plants Based on Random Amplified Polymorphic DNA(RAPD) (연초에서 발생하는 복숭아혹진딧물(Myzus persicae)형태형 2종의 Random Amplified Polymorphic DNA(RAPD)을 이용한 유전적 유연관계 분석)

  • 채순용;이기원;김상석;장영덕
    • Korean journal of applied entomology
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    • v.37 no.1
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    • pp.31-37
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    • 1998
  • Random amplified polymorphic DNA (RAPD) markers were used to analyze genetic similarity among 8 clones of apierous green peach aphid, two types (M. persicae Sulzer and M. nicotianae lack man) classified by their mo~hologi~cahla raters and host preference (Blackman, 1987), collected from tobacco plants. The genetic variation among these clones was evaluated by polymerase chain reaction amplification with 20 random primers. The higher GC contents of primers, the better in amplification efficiency of PCR reaction in general. The genetic similarities among eight aphid clones were analyzed from UPGMA (unweighted pair group average method) cluster analysis based on simple matching coefficient. The range of genetic similarity coefficients was 0.414 to 0.808. The most close relationship among the clones was similarity coefficient of 0.808 between the PG2 and the PG3 clone. The eight aphid clones analyzed were clustered into three groups by the genetic similarity coefficient. The first group, PG1, PG2, PG3 clone including in M. persicae type by their morphological characters and RED clone in M. nicotianae type was clustered at the genetic similarity coefficient of 0.643. The second group, GR1, GR2, BRN in M. nicotianae type was at the 0.636;and the third group was DBR clone in M. persicae type. The results did not indicate any correlation between m&-phological types (M. persicae and M. nicotianae) and RAPD polymorphism. We could not detect any obvious genetic relationships of the two morphological types of the green peach aphid collected from tobacco plants.

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DNA Profiles Analysed by Polymerase Chain Reaction-Random Amplified Polymorphic DNAs in Shortnecked Clam (Ruditapes philippinarum) Populations

  • Yoon, Jong-Man;Kim, Yong-Ho;Kim, Jong-Yeon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2002.08a
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    • pp.172-174
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    • 2002
  • Out of 20 primers, 6 generated a total of 1,11 major and minor RAPD bands, producing approximately 4.2 average polymorphic bands pe primer in shortnecked clam (Ruditapes philippinarum) population from Anmyeondo. The bandsharing value altered form 0.15 to 0.74, with the average of 0.5, as calculated by bandsharing analysis. The RAPD profiles obtained with DNAs of two populations from Anmyeondo and Seocheon, respectively, were considerably different (0.20 and 0.51, respectively). The varying degrees of difference among populations may also be of relevance to the retricted hybridization of wild bivalve. Besides gene mapping and breeding applications, PCR-RAPD system could be very useful for the rapid certification and quality control of seed production and for every projects based on PCR amplification of specific bivalve DNA fragments.

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Rapid molecular authentication of three medicinal plant species, Cynanchum wilfordii, Cynanchum auriculatum, and Polygonum multiflorum (Fallopia multiflorum), by the development of RAPD-derived SCAR markers and multiplex-PCR

  • Moon, Byeong-Cheol;Choo, Byung-Kil;Cheon, Myeong-Sook;Yoon, Tae-Sook;Ji, Yun-Ui;Kim, Bo-Bae;Lee, A-Young;Kim, Ho-Kyoung
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.1-7
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    • 2010
  • Definitive identification of original plant species is important for standardizing herbal medicine. The herbal medicines Cynanchi Wilfordii Radix (Baekshuoh in Korean and Beishuwu in Chinese) and Polygoni Multiflori Radix (Hashuoh in Korean and Heshuwu in Chinese) are often misidentified in the Korean herbal market due to morphological similarities and similar names. Therefore, we developed a reliable molecular marker for the identification of Cynanchi Wilfordii Radix and Polygoni Multiflori Radix. We used random amplified polymorphic DNA (RAPD) analysis of three plant species, Polygoni multiflorum, Cynanchum wilfordii, and Cynanchum auriculatum, to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed six sequence characterized amplification region (SCAR) markers for distinguishing Polygoni Multiflori Radix and Cynanchi Wilfordii Radix. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. These SCAR markers can be used for efficient and rapid authentication of these closely related species, and will be useful for preventing the distribution of adulterants.

Construction of a linkage Map in Capsicum annuum L. Using RAPD Markers and Identification of Two QTLs.

  • Yang, Tae-Jin;Kim, Yong-Jae;Park, Hyo-Guen
    • Journal of Plant Biotechnology
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    • v.1 no.2
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    • pp.109-115
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    • 1999
  • A linkage map of Capsicum annuum L. was constructed by random amplified polymorphic DNA (RAPD) markers followed in a backcross population of an intraspecific cross between cultivars HDA210 and Yatsufusa. A total of 420 random primers were tested and 311 polymorphic bands were generated by 158 random primers. Among them, 86 Yatsufusa specific bands generated by 52 primers were examined for mapping. Most bands except three segregated in Mendelian fashion fitting the expected 1:1 ratio. The total length of the map was 533 cM distributed in 15 linkage groups. The map distance between adjacent markers ranged 0 to 32.8 cM, with an average distance of 9.1 cM (63 markers). Some markers were clustered and this may be due to the amplification of a repetitive sequence by the RAPDs. Primer pairs for a sequence characterized amplified region (SCAR) were developed and the segregation scores by the SCAR primers were in accordance with the RAPD data. Two QTL markers for number of axillary shoots and for early flowering were developed. One QTL for early flowering located in the linkage group 3 and explained 61 "io of the phenotypic variation. The other QTL for the number of axillary shoots located in the linkage group 4 explained 55 % of the phenotypic variation.tion.

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Monitoring of Gentic Variability in Dicofol-susceptible, Dicofol-resistant, and its Reverse-selected Strains of Tetranychus urticae by RAPD-PCR

  • Song, Cheol;Park, Jin-Hee;Kim, Gil-Hah;Kwon, O-Yu;Cho, Kwang-Yun
    • Journal of Life Science
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    • v.9 no.1
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    • pp.14-16
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    • 1999
  • Genetic variability was monitored by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) in dicofol-susceptible (S), dicofol-resistant (R) and its reverse-selected (RS) strains of two-spotted spider mite, of Tetranychus urticae. Before the reverse-selection, RS strain, selected reversely from R strain, was 23-fold resistance ratio at {TEX}$LC_{50}${/TEX} to S strain. The resistance was started to in incline slowly to the resistance level of S strain after one year, and the resistance ratio was 4-fold in the 7 years after then. PCR-amplification of T. urticae DNA showed polymorphism in the amplifications with 12 primers in 100 kinds of arbitrary DNA sequences. RAPD amplification with primer OPR-12 (5`-ACAGGTGCGT-3`) showed amplified bands at 1,000 base pair in the S-and RS-strain, and at 350 base pair in R-strain. The results of polymorphism are genetic variabilities derived from development and selection of resistance in each strain. The peculiarly amplified fragments were guessed to participate in dicofol resistance. From the analysis of genetic similarity, it is inferred the gene composition of S-and RS-strain is much closer than that of R-strain.

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Distinction between Cold-sensitive and -tolerant Jute by DNA Polymorphisms

  • Hossain, Mohammad Belayat;Awal, Aleya;Rahman, Mohammad Aminur;Haque, Samiul;Khan, Haseena
    • BMB Reports
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    • v.36 no.5
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    • pp.427-432
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    • 2003
  • Jute is the principal coarse fiber for commercial production and use in Bangladesh. Therefore, the development of a high-yielding and environmental-stress tolerant jute variety would be beneficial for the agro economy of Bangladesh. Two molecular fingerprinting techniques, random-amplified polymorphic DNA (RAPD) and amplified-fragment length polymorphism (AFLP) were applied on six jute samples. Two of them were cold-sensitive varieties and the remaining four were cold-tolerant accessions. RAPD and AFLP fingerprints were employed to generate polymorphism between the cold-sensitive varieties and cold-tolerant accessions because of their simplicity, and also because there is no available sequence information on jute. RAPD data were obtained by using 30 arbitrary oligonucleotide primers. Five primers were found to give polymorphism between the varieties that were tested. AFLP fingerprints were generated using 25 combinations of selective-amplification primers. Eight primer combinations gave the best results with 93 polymorphic fragments, and they were able to discriminate the two cold-sensitive and four cold-tolerant jute populations. A cluster analysis, based on the RAPD and AFLP fingerprint data, showed the population-specific grouping of individuals. This information could be useful later in marker-aided selection between the cold-sensitive varieties and cold-tolerant jute accessions.

Development of PCR-Based Sequence Characterized DNA Markers for the Identification and Detection, Genetic Diversity of Didymella bryoniae with Random Amplified polymorphic DNA(RAPD)

  • Kyo, Seo-Il;Shim, Chang-Ki;Kim, Dong-Kil;Baep, Dong-Won;Lee, Seon-Chul;Kim, Hee-Kyu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.130-130
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    • 2003
  • Gummy stem blight pathogen is very difficult not only to monitor the inoculum levels prior to host infection, and also it is destructive and hard to control in field condition. We have applied RAPD technique to elucidate the genetic diversity of the genomic DNA of Didymella bryoniae and also to generate specific diagnostic DNA probe useful for identification and detection. The 40 primers produced clear bands consistently from the genomic DNA of twenty isolates of Didymella bryoniae, and two hundred seventy-three amplified fragments were produced with 40 primers. The combined data from 273 bands was analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYS-PC (Version 1.80) to generate a dendrogram. At the distance level of 0.7, two major RAPD groups were differentiated among 20 strains. RAPD group (RG) I included 8 isolates from watermelon except one isolate from melon. RAPD group (RG) IV included 12 isolates from squash, cucumber, watermelon and melon.. In amplification experiment with SCAR specific primer RG1F-RG1R resulted in a single band of 650bp fragment only for 8 isolates out of 20 isolates that should be designated as RAPD Group 1. However, same set of experiment done with RGIIF-RGIIR did not result in any amplified product.. Our attempts to detect intraspecific diversity of ITS region of rDNA by amplifying ITS region and 17s rDNA region for 20 isolates and restriction digestion of amplified fragment with 12 enzymes did not reveal polymorphic band. In order to develop RAPD markers for RGIV specific primer, a candidate PCR fragment( ≒1.4kb) was purified and Southern hybridized to the amplified fragment RGIV isolates. This promising candidate probe recognized only RGIV isolates

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Evaluation of Genetic Diversity among the Genus Viola by RAPD Markers

  • Oh, Boung-Jun;Ko, Moon-Kyung;Lee, Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.19 no.6
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    • pp.716-720
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    • 2006
  • The genetic diversity among the genus Viola was evaluated using the random amplified polymorphic DNA (RAPD) method. A total of 142 distinct amplification fragments by 18 random primers were scored to perform the cluster analysis with UPGMA. Viola species from the subsection Patellares were clustered into group I to IV. The groups from I to IV were consistent with its morphological taxonomy, series Pinnatae, Chinensis, Variegatae, and Patellares in the subsection Patellares, respectively. Even though V. albida and V. albida var. takahasii were classified in Chinensis, they were assigned into group I. The cluster analysis separated other subsections from Patellares in the section Nomimium. Interestingly, V. verecunda and V. grypoceras in subsections Biobatae and Trigonocarpae, respectively, were clustered into group C with a high similarity coefficient. Therefore, RAPD analysis can be used for providing an alternative classification system to identify genotypes and morphological characters of Viola species.

Evaluation of Genetic Diversity among Korean Wild Codonopsis lanceolata by Using RAPD

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    • Korean Journal of Plant Resources
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    • v.10 no.3
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    • pp.258-264
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    • 1997
  • The introduction of molecular biology methodologies to plant improvement programs offers an invaluable opportunity for extensive germplasm characterization. We have applied the developed technique of random amplification of polymorphic DNA(RAPD)to the analysis of evaluating genetic diversity among Korean wild Codonopsis lanceolata. A total of 340 polymorpic hands were gernerated on agarose- and polyacrylamide-gel by 19 primers of abitrary sequence. grouped by cluster analysis using sample matching coefficients of similarity. Among of the samples. the minimum genetic distance value was obtained between sample no. 1(Girisan) and no. 2(Girisan), and the largest value between sample no. 11(Sulaksan) and no. 17(Sulaksan).In separate cluster dendrograms based on agareose - and polyacryamide-gel. some differences were observed; In the case of agarose gel,41 samples could be devided into 7 groups at below about 0.44 level of distance. However they were divided into 6 gourps at below about 0.40 level of distance in the case of polyacrylamide gel. These results showed that polymophic data in agrose were not grouped to wild plant selected from each mountainous district except for wild plants selected from Sulaksan and Chiaksan. We believe that polyacrylamide-RAPD is a superior method for detecting DNA polymorphism compared to agarose-RAPD method.

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