• Title/Summary/Keyword: polymorphic DNA

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Genetic relationships and molecular authentication of plant origins and the commercial medicinal herbs in peony using RAPD markers

  • Bang, Kyong-Hwan;Jung, Jin-Ho;Kim, Ok-Tae;Chung, Jong-Wook;Ham, In-Hye;Seong, Nak-Sul;Luo, Rong;Zhang, Gui-Jun;Choi, Ho-Young
    • Advances in Traditional Medicine
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    • v.7 no.1
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    • pp.26-33
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    • 2007
  • Genetic polymorphism and molecular authentication were investigated with the commercial medicinal herb, Peony (Paeonia spp.), using random amplified polymorphic DNA (RAPD) markers. To identify the polymorphism of the RAPD patterns among plant origins, 20 different random primers were applied to the genomic DNA extracted from Paeonia spp. plants such as Paeonia (P.) lactiflora, P. officinale and P. japonica. Ten primers out of 20 primers could be used to discriminate the plant species in the same genus and 72 out of 81 scored DNA fragments (88.9%) generated with these primers were polymorphic. Especially, four primers, such as OPA1, OPA3, OP9, and OPA13, were useful to discriminate the plant origins among the species of Peony. In the results of cluster analysis using RAPD data obtained from the 10 primers, Peony (Paeonia spp.) plants used in this study were grouped into the two distinctive clusters, genetically. Herb medicine, especially P. lactiflora, were easily identified, when species-specific primers were applied to the investigation for discriminating herb medicine currently traded in domestic herb market, Kyungdongmart. Consequently, RAPD analysis was useful method to discriminate plant origins and the commercial medicinal herbs, Paeonia spp..

Mycelial Propagation and Molecular Phylogenetic Relationships of Commercially Cultivated Agrocybe cylindracea based on ITS Sequences and RAPD

  • Alam, Nuhu;Kim, Jeong-Hwa;Shim, Mi-Ja;Lee, U-Youn;Lee, Tae-Soo
    • Mycobiology
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    • v.38 no.2
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    • pp.89-96
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    • 2010
  • This study evaluated the optimal vegetative growth conditions and molecular phylogenetic relationships of eleven strains of Agrocybe cylindracea collected from different ecological regions of Korea, China and Taiwan. The optimal temperature and pH for mycelial growth were observed at $25^{\circ}C$ and 6. Potato dextrose agar and Hennerberg were the favorable media for vegetative growth, whereas glucose tryptone was unfavorable. Dextrin, maltose, and fructose were the most effective carbon sources. The most suitable nitrogen sources were arginine and glycine, whereas methionine, alanine, histidine, and urea were least effective for the mycelial propagation of A. cylindracea. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The sequence of ITS2 was more variable than that of ITS1, while the 5.8S sequences were identical. The reciprocal homologies of the ITS sequences ranged from 98 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) using 20 arbitrary primers. Fifteen primers efficiently amplified the genomic DNA. The average number of polymorphic bands observed per primer was 3.8. The numbers of amplified bands varied based on the primers and strains, with polymorphic fragments ranging from 0.1 to 2.9 kb. The results of RAPD analysis were similar to the ITS region sequences. The results revealed that RAPD and ITS techniques were well suited for detecting the genetic diversity of all A. cylindracea strains tested.

Effect of Sequence Variation in Bovine Mitochondrial DNA D-loop Region on Economic Traits for Hanwoo (한우 경제형질에 미치는 Mitochondrial DNA D-loop 영역의 염기서열 변이효과)

  • Oh, J.D.;Yoon, D.H.;Kong, H.S.;Lim, H.J.;Lee, H.K.;Cho, B.W.;Hong, K.C.;Jeon, G.J.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.933-938
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    • 2003
  • This study was performed to analyse the sequences of variations of mtDNA D-loop and their effects on carcass traits in Hnawoo(Korean cattle). The resulting sequences were compared with previously published sequences for other cattle breeds(GenBank J01394). The PCR was used to amplify a total of 964 bp between nucleotide 15758 and 383 within D-loop region of mtDNA using specific primers. Twenty five polymorphic sites by nucleotide substitution were found in mtDNA of Hanwoo. The frequencies of positions at 169, 16042, 16093, 16119, 16255 and 16302 nt with high levels of sequence polymorphism were 0.891, 0.117, 0.109, 0.182, 0.197 and 0.117, respectively. The substitution effect at 169 and 16119 nt was found significant on marbling score. Also substitution effect at 169 and 16042 nt was highly significant(p〈0.01) on backfat. thickness. Polymorphism of mtDNA sequence in D-loop region could be useful for the analysis of cytoplasmic genetic variation and associations with the other economically important traits and maternal lineage analysis in Hanwoo.

Effect of Sequence Variation in Mitochondrial DNA D-loop Region on Milk and Milk Fat Production in Holstein Cows (Holstein의 유량과 유지방 생산에 미치는 Mitochondrial DNA D-loop 영역의 염기 서열 변이 효과)

  • Oh J. D.;Kong H. S.;Lee H. K.;Jeon G. J.
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.9-13
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    • 2005
  • This study was performed to analyze the sequence variation in mtDNA D-loop and their effects on milk and milk fat production in Holstein cows. The analyzed sequences were compared with previously published sequences from other cattle breeds (GenBank J01394). PCR was performed to amplify a total of 964 bp between nucleotide 15758 and 383 within D-loop region of mtDNA using specific primers. Thirty five polymorphic sites by nucleotide substitution were found in mtDNA. The frequencies of positions at 106, 169, 16057, 16231 and 16255 nt with high levels of sequence polymorphism were 0.090, 0.555, 0.055, 0.090 and 0.050, respectively. The substitution effect at 169 nt was found significant on milk production, and substitution effect at 16118, 16139 and 16302 nt was highly significant (p<0.1) on milk fat production. Polymorphism of mtDNA sequence in D-loop region might be useful for the analysis of cytoplasmic genetic variation and associations with the other economically important traits and maternal lineage analysis in Holstein cows.

Potential Biotypes in Korean Isolates of Bipolaris cactivora Associated with Stem Rot of Cactus

  • Kim, Jeong-Ho;Jeoung, Myoung-Il;Hyun, Ick-Hwa;Kim, Young-Ho
    • The Plant Pathology Journal
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    • v.20 no.3
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    • pp.165-171
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    • 2004
  • A total of 62 isolates of Bipolaris cactivora causing cactus stem rots were isolated from major cactus-growing areas in Korea. Colony morphology of the isolates on potato-dextrose agar was differentiated into aerial (CA) and non-aerial mycelial types (CB). CA had profound aerial mycelium with grayish brown (CA-l), light brownish (CA-2), and brownish (CA-3) pigmentations; respectively, while CB had dark brownish pigmentations. CA had conidia of less dark pigmentation and acute terminal end. CB had darker and more round-end conidia. Twenty-eight amplified fragments were produced by polymerase chain reaction (PCR) with a set of 2 random primers. The sizes of amplified DNA fragments ranged approximately from 0.1 to 2.3 kb. The isolates were classified into 2 major genomic DNA random amplified polymorphic DNA (RAPD) groups at the genomic similarity of 97.7% and 95.1%, respectively. Cluster analysis of genetic similarity among the isolates generated a dendrogram that clearly separated all isolates into SA or SB. This result suggests that there may be two morphotypes of B. cactivora in Korea that may differ in their genetic constitutes.

Molecular Identification Patterns of Clinical Isolates from Korean Patients Infected with Dermatopbytes

  • Kim, Sang-Ha;Cho, Seong-Wan;Hwang, Seock-Yeon;Jeon, Soo-Young;Kim, Young-Kwon
    • Biomedical Science Letters
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    • v.16 no.3
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    • pp.187-192
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    • 2010
  • The aim of this study was to compare the results of morphological phenotypic examination and molecular biological method, randomly amplified polymorphic DNA (RAPD) using clinical isolates from Korean patients infected with dermatophytes. A total of 44 clinical isolates have been collected by Korean Collection Medical Fungi (KCMF) and those were subjected for the RAPD analysis. The results showed that the fungal strains that have RG1 band patterns were identified as Trichophyton rubrum by the neighbour-joining analysis. Additionally, other fungal strains that have RG3 band patterns were identified as Epidermophyton floccosum and RG5 band patterns were identified as Micorsporum gypseum. Even though molecular biological method such as RAPD are not necessary, it might be useful when some strains have similar morphological characteristics or when same species has phenotypic variations. In this study, therefore, we targeted internal transcribed spacer (ITS) regions of fungal rDNA, performed RAPD and then compared with morphological phenotypic examination.

Taxonomic Position of Korean Isolates of Rhizoctonia solani Based on RAPD and ITS Sequencing of Ribosomal DNA

  • Jeon, Young-Ah;Kim, Wan-Gyu;Kim, Dae-Ho;Kwon, Soon-Wo;Hong, Seung-Beom
    • The Plant Pathology Journal
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    • v.26 no.1
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    • pp.83-89
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    • 2010
  • Taxonomic position of 46 Korean isolates of Rhizoctonia solani which were classified into nine intraspecific groups by anastomosis and cultural characteristics was analyzed by randomly amplified polymorphic DNA (RAPD) and sequence analyses of the internal transcribed spacer (ITS) regions of ribosomal DNA. All the isolates within each group showed highly similar band patterns in RAPD. The ITS regions of the isolates within the same groups showed a high level of sequence similarity above 96.0% whereas similarities among different groups were below 94.4%. When compared with several reference strains of R. solani from foreign countries, all the Korean isolates were clustered with the foreign isolates belonging to the same groups in the phylogenetic tree. All six Korean strains of AG-4 were identified as HG-1 out of 3 subgroup of AG-4. We discussed taxonomic position of Korean isolates of R. solani and showed that sequence analysis with ITS regions could be a rapid and useful method for identification of intraspecific group of R. solani.

Genetic Linkage Mapping of RAPD Markers Segregating in Korean Ogol Chicken - White Leghorn Backcross Population

  • Hwang, K.C.;Song, K.D.;Kim, T.H.;Jeong, D.K.;Sohn, S.H.;Lillehoj, H.S.;Han, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.3
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    • pp.302-306
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    • 2001
  • This study was carried out to construct mapping population and to evaluate the methods involved, including polymorphic DNA marker system and appropriate statistical analysis. As an initial step to establish chicken genome mapping project, White Leghorn (WL) and Korean Ogol chicken (KOC) were used for generating backcross population. From 8 initial parents, total 280 backcross progenies were obtained and 40 were used for genotyping and linkage analysis. For development of novel polymorphic markers for KOC, Random Amplified Polymorphic DNA (RAPD) markers specific for this chicken line were generated. Also included in this study were six microsatellite markers from East Lansing map as reference loci. For segregation analysis, 15 RAPD markers and 6 microsatellites were used to genotype the backcross population. Among the RAPD markers that we developed, 2 pairs of markers were identified to be linked and another 4 RAPD markers showed linkage with microsatellites of known map. In summary, this study showed that our backcross population generated from the mating of KOC to WL serves as a valuable genetic resource for genotyping. Furthermore, RAPD markers are proved to be valuable in linkage mapping analysis.

Studies on Genetic Variation of Different Chinese Duck Populations with Random Amplified Polymorphic DNA Analysis

  • Su, Y.;Liu, C.W.;Liu, L.;Ye, C.H.;Cao, W.Q.;Huang, Y.Q.;Zheng, J.;Cai, D.Y.;Olowofeso, O.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.4
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    • pp.475-481
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    • 2006
  • The genetic polymorphism and relationships of Muscovy, Cherry Valley Meat ducks, Partridge ducks and their crossbreds $F_1$ and $F_2$, respectively, were studied using a random amplified polymorphic DNA (RAPD) technique. The results showed that RAPD markers were effective for the analysis of genetic relationships among ducks. Amplification with 20-primers gave 760 reproducible amplified fragments. The percentage of polymorphic marker band was 74.70%, which indicates that the RAPD technique had higher efficiency of polymorphism detection and sensitivity in studying the genetic variations among ducks and showed that the genetic polymorphism was abundant between two species of duck populations. The average index of genetic distance in hybrid $F_2$ was 0.2341 and higher than that of its parents, which indicates that the genetic diversity was improved by crossbreeding with Muscovy.

Genetic Diversity and Population Structure of Comus controversa Hemsley Using RAPD (RAPD에 의한 층층나무의 유전적 다양성과 집단구조)

  • Moon, Sung-Gi;Huh, Man-Kyu
    • Journal of Life Science
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    • v.18 no.2
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    • pp.175-179
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    • 2008
  • Cornus controversa is a long-lived woody species mostly distributed in East Asia. Random amplified polymorphic DNA (RAPD) markers were used to investigate the genetic diversity and population structure of Korean populations of this species. A high level of genetic variation was found in seven populations of C. controversa. The mean genetic diversity (H) was 0.222 across populations, varying from 0.200 to 0.238. Eighty of the 93 loci (86.0%) showed detectable polymorphism in at least one population. Total genetic diversity values ($H_T$) varied between 0.192 and 0.231, giving an average overall polymorphic loci of 0.212. The interlocus variation of genetic diversity within populations ($H_S$) was high (0.167). Mean of genetic diversity in C. controversa was higher than average values for species with similar life history traits. The sexual reproduction, perennial habitat, and longevity are proposed as possible factors contributing to high genetic diversity. On a per locus basis, the proportion of total genetic variation due to differences among populations ($G_{ST}$) ranged from 0.169 to 0.278 with a mean of 0.216, indicating that about 21.6% of the total genetic variation was among populations. An indirect estimate of the number of migrants per generation (Nm=1.893) indicated that gene flow was extensive among Korean populations of C. controversa.