• Title/Summary/Keyword: Random amplified polymorphic DNA (RAPD)

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Phylogenetic Relationship of Ligularia Species Based on RAPD and ITS Sequences Analyses (RAPD 및 ITS 염기서열 분석을 이용한 곰취 속(Ligularia) 식물의 유연관계 분석)

  • Ahn, Soon-Young;Cho, Kwang-Soo;Yoo, Ki-Oug;Suh, Jong-Taek
    • Horticultural Science & Technology
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    • v.28 no.4
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    • pp.638-647
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    • 2010
  • The genetic relationships in 5 species of $Ligularia$ were investigated using RAPD (Randomly Amplified Polymorphic DNA) and ITS (Internal Transcribed Spacer) sequences analyses. In RAPD analysis, sixty three of 196 arbitrary primers showed polymorphism. The amplified fragments ranged from 0.2 to 1.6 kb in size. The dendrogram was constructed by the UPGMA clustering algorithm based on genetic similarity of RAPD markers. A total of 16 accessions were classified into 5 major groups corresponding each species at the similarity coefficient value of 0.77. In the ITS sequence analysis, the size of ITS 1 was varied from 248 to 256 bp, while ITS 2 was varied from 220 to 222 bp. The 5.8S coding region was 164 bp in lengths. Forty nine sites (10.2%) of the 478 nucleotides were variable, and the G+C content of ITS region ranged from 49.4 to 53.5%. In the ITS tree, five species of $Ligularia$ were monophyletic, and $L.$ $taquetii$ was the first branching within the clade. $Ligularia$ $intermedia$ formed a clade with $L.$ $fischeri$ var. $spiciformis$ (BS=79), and $L.$ $stenocephala$ and $L.$ $fischeri$ were also claded. Two data sets were congruent, except of the position of $L.$ $fischeri$ var. $spiciformis$.

Morphological and Genetic Diversity of Korean Native and Introduced Safflower Germplasm

  • Shim Kang-Bo;Bae Seok-Bok;Lim Si-Kyu;Suh Duck-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.4
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    • pp.337-341
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    • 2004
  • Morphological and genetic diversity of thirty nine safflower germplasm were collected and evaluated by Principal Component Analysis (PCA) and Random Amplified Polymorphic DNA (RAPD) method. Stem length and seeding to flowering days of the safflower germplasm showed $26\~117cm\;and\;76\~179$ days of variation respectively. USA originated germplasm showed higher oil content as $39\%$, but that of Japanese showed lower as $26\%$. PCA made three different cluster groups according to some agronomic characteristics of safflower. Korea originated germplasm showed similar cluster group with that of collected from USA in the PCA of stem length. But in the seeding to flowering days, it showed similar cluster pattern with that of collected from Japan rather than USA. In the experiment of RAPD analysis, total five primers showed polymorphism at the several chromosomal loci. Korea, China Japan and South Central Asia originated germplasm were differently classified with USA and South West Asia originated germplasm with lower similarity coefficient value (0.47). Most of Korea originated germplasm were grouped with South Central Asia originated germplasm with higher similarity coefficient value (0.74) conferring similar genetic background between both of them. China and Japan originated germplasm were dendrogramed with Korea originated germplasm at the 0.65 and 0.50 similarity coefficient values respectively. Some common results were expected from both of PCA and RAPD analysis, but lower genetic heritability caused by relative higher portion of environmental variance and environment by genotype interaction at the expression of those of agronomic characteristics made constraint to find any reliable results.

Diversity of the Streptococcal Strains Isolated from Diseased Olive Flounder (Paralichthys olivaceus) (넙치 (Paralichthys olivaceus) 병어에서 분리된 연쇄상구균의 다양성)

  • KIM Jong-Hun;KIM Eunheui
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.6
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    • pp.654-660
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    • 2003
  • To evaluate the biological diversity of fish pathogenic streptococci, 35 strains isolated from diseased olive flounder (Paralichtys olivaceus), were analyzed using a random amplified polymorphic DNA (RAPD) technique with the oligonucleotide commercial primer 6 (Amersham Biosciences). Api 20 Strep test, drug resistance and artificial infection were carried out for further characterization of the isolates. RAPD fingerprints showed similar pattern in 25 strains (about $71.4\%$ of 35 isolates) and these strains were designed as RA group 1. Similarities greater than $44\%$ were obtained when the Dice coefficient was applied among the isolates of RA 1. On the other hand, the reference Streptococcus iniae showed a similar RAPD profile to the isolates with similarity levels of $40-93.3\%.$ Rh I was suggested to be the dominant group isolated from olive flounder suffering from streptococcosis. However, the isolates of Rh 1 group were not classified into the same species by the Api 20 Strep identification system. There was no peculiarity in drug resistance patterns of Rh I group isolates against 7 antibacterial agents. However, only 3 of 25 isolates $(0.12\%)$ showed oxytetracycline (OTC) resistance and OTC might be a useful chemotherapeutic agent in controlling the streptococcosis by strains of RA I group in olive flounder. Fish injected intraperitoneally with $10^5$ CFU of an isolate of Rh I and RA III group showed $60\%\;and\;50\%$ accumulative mortality for 20 days, respectively ($20\%$ in control or Rh II). However luther comparative studies about differences in virulence between isolates are needed.

Systematic Relationships among Species of the Genus Dendronephthya (Alcyonacea: Octocorallia; Anthozoa) Based on RAPD Analysis

  • Song, Jun-Im;Lee, Young-Ja
    • Animal cells and systems
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    • v.4 no.1
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    • pp.1-7
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    • 2000
  • The genus Dendronephthya, generally known as soft corals, is reported as an abundant and variable taxon. They mostly distribute in warmer waters of the Undo-Pacific Ocean region including Korea. In spite of their abundance and ecological importance as habitats of marine organisms, there are difficulties in the study of their identification and systematics because they have morphological variabilities and limited taxonomec characters. To resolve the problems, we attempted to elucidate the genetic relationships in the genus Dendronephthya by using random amplified polymorphic DNA (RAPD) analysis. This study was based on eight dendronephthian species and one Alcyoniidae species, Alcyonium gracillimum, as an outgroup. The results from all analysis suggest that they could be classified into four groups by the growth form and the anthocodial grades as follows: the first one,D. putteri and D. suensoni with the divaricate form and VI grade; the second one,D. sp.1 and D. sp.2 with the divaricate form and III or IV grade; the third one, D. gigantea and D. aurea being closer than D. spinifera with the glomerate form and III grade; the last one, D. castanea related to D. gigantea rather than D. putteri with the umbellate form and IV grade. Moreover, the divaricate form was separated from the group of the glomerate and umbellate form. At the intraspecies level, the types of the D. castanea, D. gigantea and D. spinifera were separated depending on the feature of spicules in the polyp head, and the coloration could not influence genetic variation. From this study, we can confirm that their morphological characters are compatible with the genetic variation, also RAPD analysis is a very useful method for resolving the systematic relationships of den-deonephthians.

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Analysis of Genetic Relationship among Korean Native Orostachys Species Using RAPD (RAPD를 이용한 자생 바위솔속(Orostachys) 식물의 유연관계 분석)

  • Lee, Byoung Ae;Kim, Hag Hyun;Cho, Yong Gu;Lee, Cheol Hee
    • Horticultural Science & Technology
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    • v.19 no.2
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    • pp.159-162
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    • 2001
  • The genetic relationships of Korean native Orostachys species collected from various regions were analysed using random amplified polymorphic DNA (RAPD) method. Eighteen Orostachys species analysed with UPGMA were clustered into three groups A, B, and C. However, four species were not clustered into any group. O. iwarenge species in group A (No. 18-No. 22) showed low similarity with 66.3-73.9% according to the cluster analysis. O. malacophyllus species in group B (No. 12-No. 17) showed low similarity with 66.7-83.7% according to the cluster analysis. The similarity coefficient value of O. japonicus (No. 3-No. 8) except Anmyeondo collected variety (No. 9) showed higher level with 84.2-92.3% than O. iwarenge or O. malacophyllus. Therefore, O. japonicus is thought to be genetically stable, and have less regional variation.

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Resistance Potential of Bread Wheat Genotypes Against Yellow Rust Disease Under Egyptian Climate

  • Mahmoud, Amer F.;Hassan, Mohamed I.;Amein, Karam A.
    • The Plant Pathology Journal
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    • v.31 no.4
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    • pp.402-413
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    • 2015
  • Yellow rust (stripe rust), caused by Puccinia striiformis f. sp. tritici, is one of the most destructive foliar diseases of wheat in Egypt and worldwide. In order to identify wheat genotypes resistant to yellow rust and develop molecular markers associated with the resistance, fifty F8 recombinant inbred lines (RILs) derived from a cross between resistant and susceptible bread wheat landraces were obtained. Artificial infection of Puccinia striiformis was performed under greenhouse conditions during two growing seasons and relative resistance index (RRI) was calculated. Two Egyptian bread wheat cultivars i.e. Giza-168 (resistant) and Sakha-69 (susceptible) were also evaluated. RRI values of two-year trial showed that 10 RILs responded with RRI value >6 <9 with an average of 7.29, which exceeded the Egyptian bread wheat cultivar Giza-168 (5.58). Thirty three RILs were included among the acceptable range having RRI value >2 <6. However, only 7 RILs showed RRI value <2. Five RILs expressed hypersensitive type of resistance (R) against the pathogen and showed the lowest Average Coefficient of Infection (ACI). Bulked segregant analysis (BSA) with eight simple sequence repeat (SSR), eight sequence-related amplified polymorphism (SRAP) and sixteen random amplified polymorphic DNA (RAPD) markers revealed that three SSR, three SRAP and six RAPD markers were found to be associated with the resistance to yellow rust. However, further molecular analyses would be performed to confirm markers associated with the resistance and suitable for marker-assisted selection. Resistant RILs identified in the study could be efficiently used to improve the resistance to yellow rust in wheat.

Intergeneric Transfer of Isolated Nuclei from Lentinus edodes into Protoplasts of Pleurotus florida (사철느타리버섯 원형질체내에 표고 핵의 전이)

  • Yoo, Young-Bok;Shin, Pyung-Gyun
    • The Korean Journal of Mycology
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    • v.24 no.2 s.77
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    • pp.111-126
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    • 1996
  • Transfer of the isolated nuclei from Lentinus edodes into protoplasts of Pleurotus florida was induced with polyethlene glycol (PEG) and $CaCl_2$. The intergeneric transfer products were classified into nuclear hybrid, heterokaryon or synkaryon, and reconstituted cell. These progenies except nuclear hybrids formed mature fruiting bodies on sawdust rice bran medium. Formation of fruit bodies was influenced by several factors such as light, temperature, nutrition and physic state of the culture media. Most of fruiting body characters were similar to those of P. florida in synkaryon and L. edodes in reconstituted cell, respectively. All these basidiocarps had clamp connections though initial heterokaryon colonies were lacking. Isozyme patterns of intergeneric progenies were quite different from those of parents. DNA polymorphisms of transfer products were also compared by random amplified polymorphic DNAs (RAPD) analysis based on polymerase chain reaction. The RAPD patterns were different from those of donor and recipient. DNA fingerprints ranged in size from 0.25 to 4.0 Kb. On the basis of RAPD, the transfer products were classified into five groups. Two synkaryon were analysed with distribution of progenies and segregation of genetic markers by random spore analyses. The genetic markers were segregated into wild type and riboflavine requiring auxotrophs.

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The classification and comparison of genetic diversity of genus Malus using RAPD (RAPD를 이용한 능금속 식물종의 계통관계와 유전적 다양성)

  • Huh, Man-Kyu
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.756-761
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    • 2007
  • Cenus Malus is a long-lived woody species primarily distributed throughout Asia. Many species of this genus are regarded as agriculturally and ecologically important. The phynetics and genetic diversity among eight species of genus Malus were reconstructed using the random amplified polymorphic DNA (RAPD) markers. In a simple measure of intraspecies variability by the percentage of polymorphic bands, the M. micromalus exhibited the lowest variation (34.7%). The M. pumila showed the highest (50.0%). Mean number of alleles per locus (A) ranged from 1.347 to 1.500 with a mean of 1.437. The phenotypic frequency of each band was calculated and used in estimating genetic diversify (H) within species. The mean of H was 0.190 across species, varying from 0.155 to 0.220. In particular, two cultivated species, M. pumila and M. asiatica, had high expected diversity, 0.314 and 0.307, respectively. On a per locus basis, the proportion of total genetic variation due to differences among species ranged from 0.388 to 0.472 with a mean of 0.423, indicating that 42.3% of the total variation was found among species. The phylogenetic tree showed three distinct elates. One includes M. sieversii, M. pumila, and M. asiatica. Another includes three M. baccata taxa. The other includes M. sieboldii, M. floribunsa, and M. micromalus. One variety and one form of M. sieboldii were well separated each other. RAPD markers are useful in germ-plasm classification of genus Malus and evolutionary studies.

An analysis of the genetic diversity of a riparian marginal species, Aristolochia contorta (수변 경계종인 쥐방울덩굴의 유전적 다양성 분석)

  • Nam, Bo Eun;Park, Hyun Jun;Son, Ga Yeon;Kim, Jae Geun
    • Journal of Wetlands Research
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    • v.22 no.2
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    • pp.100-105
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    • 2020
  • Northern pipevine (Aristolochia contorta) commonly inhabits marginal areas between waterside and terrestrial vegetation. In particular, A. contorta is ecologically important in the marginal areas as a food plant of dragon swallowtail butterfly (Sericinus montela), which is designated as vulnerable species in the Republic of Korea. For long-term sustainability of the plant population, assessment of the genetic diversity of exist populations should be conducted. Genomic DNA of A. contorta leaf samples were extracted from four populations where the vigorous growth were observed in the South Korea. Intra-population genetic diversity and inter-population genetic distance were assessed using randomly amplified polymorphic DNA (RAPD) with five polymorphic random primers. Overall genetic diversity was lower, compared to other wetland species (h: 0.0607 ~ 0.1401; I: 0.0819 ~ 0.1759), while GP showed the highest intra-population genetic diversity. Despite of the geographical distance, GP showed the larger genetic distance from other populations. This result seemed to be caused by the fragmented habitat and lower sexual reproduction of A. controta. Mixture of the different source populations and construction of the proper environmental condition such as shade and physical support for sexual reproduction should be considered for conservation of A. contorta population.

Molecular Typing of Staphylococcus aureus Strains from Domestic Animals and Humans by REP-PCR Analysis (REP-PCR을 이용한 국내 사람과 동물유래 Staphylococcus aureus 분리주의 Molecular Typing)

  • Woo Yong-Ku;Kim Shin
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.60-66
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    • 2005
  • To select the rapid and efficient molecular subtyping method for epidemiologic monitoring of Staphylococcus aureus (S. aureus) strains at clinical laboratory levels, a total 116 of S. aureus and MRSA (methicillin-resistant S. aureus) strains from diverse animal species [Korean cattle, goat, pig, dog, chicken, mouse] and also humans were analyzed. To evaluate the discriminatory ability (DA) of individual PCR methods, random amplified polymorphic of DNA [RAPD; 4M & RA primer], repetitive extragenic palindromic sequences PCR (REP-PCR), and enterobacterial repetitive intergenic consensus sequences PCR (ERIC-PCR) methods were conducted and then compared on their Simpson's index of diversity (SID) values based on the dendrogram patterns, which was produced by software program (BiolD2+ & GelCompar II). In first, RAPD using the 4M primer (SID 0.915) was expressed more higher SID value than that of RA primer (SID 0.874). 4M primer was expressed more powerful DA than RA. Both REP-PCR (SID 0.930) and ERIC-PCR (SID 0.929) methods showed much more higher DA than that of RAPD. According to the present results, both REP-PCR and ERIC-PCR among the tested analysis methods were found as the most reliable and discriminative molecular subtyping method, because they expressed the highest DA for the present S. aureus and MRSA strains.