• Title/Summary/Keyword: RAPD primer

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Discrimination of Potato Varieties by Random Amplified Polymorphic DNA Analysis (RAPD에 의한 감자 품종의 구분)

  • Seo, Hyo Won;Yi, Jung Yoon;Cho, Hyun Mook;Park, Young Eun;Oh, Seung Eun
    • Horticultural Science & Technology
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    • v.19 no.1
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    • pp.29-33
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    • 2001
  • This study was carried out to discriminate potato cultivars and breeding lines by specific molecular markers using random amplified polymorphic DNA (RAPD) analysis. The genotypes of potatoes used for analysis were eight cultivars and five breeding lines. Some of those show much phenotypic resemblances among them because 'Jopung', 'Daekwan70', 'Gawon', and 'Daekwan72' have immediate parental relationship with 'Superior', 'Irish Cobbler', 'Namsuh', and 'Atlantic', respectively. So, there are many difficulties to distinguish the varieties by the morphological characteristics. Three URP primers, URP2, URP4, and URP8 were selected for promising primers to discriminate potato genotypes or cultivars. The three URP primers were shown very high reproducibility because of the relatively high annealing temperature and long primer size. Although the results of similarity analyses did not always reflect the genetic relationship between potato varieties, the reproducible pattern of amplified DNA bands by URP primers showed possibility for molecular markers for discrimination of potato genotype or cultivar.

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Specific and Sensitive Detection of Venturia nashicola, the Scab Fungus of Asian Pears, by Nested PCR

  • Koh, Hyun Seok;Sohn, San Ho;Lee, Young Sun;Koh, Young Jin;Song, Jang Hoon;Jung, Jae Sung
    • The Plant Pathology Journal
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    • v.29 no.4
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    • pp.357-363
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    • 2013
  • The fungus Venturia nashicola is the causal agent of scab on Asian pears. For the rapid and reliable identification as well as sensitive detection of V. nashicola, a PCR-based technique was developed. DNA fingerprints of three closely related species, V. nashicola, V. pirina, and V. inaequalis, were obtained by random amplified polymorphic DNA (RAPD) analysis. Two RAPD markers specific to V. nashicola were identified by PCR, after which two pairs of sequence characterized amplified region (SCAR) primers were designed from the nucleotide sequences of the markers. The SCAR primer pairs, designated as D12F/D12R and E11F/E11R, amplified 535-bp and 525-bp DNA fragments, respectively, only from genomic DNA of V. nashicola. The specificity of the primer sets was tested on strains representing three species of Venturia and 20 fungal plant pathogens. The nested PCR primer pair specific to V. nashicola was developed based on the sequence of the species-specific 525-bp DNA fragment amplified by primer set E11F/E11R. The internal primer pair Na11F/Na11R amplified a 235-bp fragment from V. nashicola, but not from any other fungal species tested. The nested PCR assay was sensitive enough to detect the specific fragment in 50 fg of V. nashicola DNA.

Analysis of Genetic Diversity in Soybean Varieties Using RAPD Markers (사료작물로 이용이 가능한 한국 재배콩의 RAPD 표지인자에 의한 유전적 다양성 분석)

  • Lee, Sung-Kyu;Kim, Bum-Jun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.18 no.4
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    • pp.277-284
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    • 1998
  • Random amplified polymorphic DNA (RAPD) analysis was used to detect the genetic diversity of soybean (Glycine max (L.) Merr.) varieties and field bean (Glycine soza Sieb. and Zucc.) Five soybean varieties and one field bean were analysed with random primers using the polymerase chain reaction (PCR). Nine primers of a total twenty random primer were selected to amplify DNA segments. A total of 74 PCR products were amplified and 67.6% of which were polymorphic. The size of DNA molecule is ranged 0.13~2.0Kb and typically generated four to eight major bands. Specific genetic marker were revealed in primer sequence 5'-CAG GCC CIT C-3', 5'-TGC TCT GCC C-3' and 5'-GTC CAC ACG G-3', respectively. Genetic similarity between each of the varieties were calculated from the pair-wise comparisons of amplification products and a dendrogram was constructed by an unweighted pair-group method with arithmethical means (UPGMA). The results indicate that intervarietal relationships of soybean have a narrow genetic base and between the varieties, Hwanggum-kong and Seckryang-bootkong is more closely related than the rest of varieties, and field bean is related quite distant.

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Analysis of antibiotic susceptibility of Salmonella Enteritidis isolated from Gyeongnam province and the bacterial genotyping by using RAPD-PCR (경남지역에서 분리한 Salmonella Enteritidis의 항생제 감수성 검사 및 random amplification polymorphic DNA (RAPD)-PCR을 이용한 유전형 분석)

  • Kim, Eun-Gyeong;Kim, Min-Kyung;Kwon, Hyun-Ae;Youn, Do-Kyung;Koo, Jeong-Heon;Park, So-Yeon;Lee, Hui-Geun;Jo, Myeong-Hui;Hah, Do-Yun;Kim, Cheol-Ho;Hwang, Bo-Won;Kim, Sang-Hyun
    • Korean Journal of Veterinary Service
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    • v.41 no.3
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    • pp.149-155
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    • 2018
  • Salmonella Enteritidis (S. Enteritidis) are found in animals, humans, and environment. In addition, S. Enteritidis draws attention to the public health concerns due to carriage of antibiotic resistance traits. For these reasons, the prevalence and antibiotic resistance patterns of S. Enteritidis are significant issues with regard to public health. To address this issues, a total of 24 strains of S. Enteritidis from 164 samples collected from several slaughterhouses in Gyeong-Nam province in order for antibiotic resistance profiles. Subsequently, we characterized the genotyping by random amplification polymorphic DNA (RAPD)-PCR. As a result, very high level of resistance to protein synthesis inhibition antibiotics and most isolates were susceptible to others. Six random primers were used for RAPD-PCR to reveal genotypes of S. Enteritidis isolates. One of the primer, P1245, generated 147 distinct RAPD-PCR fragments ranging from 400~3000 bp. The number of RAPD-PCR products ranged from 4 to 8 for this primer. The RAPD-PCR fragments could be placed these strains into 3 subgroups and 2 classes by UPGMA cluster analysis. Interestingly, several S. Enteritidis that isolated from different slaughterhouses showed same genotype. These results showed only limited genetic variation among the isolates, those were grouped into a few different patterns of antibiotic resistance.

Phylogenetic Relationships and Genetic Diversity in Collected Resources of Carthamus tinctorius by Random Amplified Polymorphic DNA Markers (RAPD 마커에 의한 수집된 홍화자원에서 계통관계와 유전적 다양성)

  • Sung, Jung-Sook;Cho, Gyu-Taek;Lee, Gi-An;Baek, Hyung-Jin;Huh, Man-Kyu
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1764-1771
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    • 2010
  • Carthamus tinctorius L. (Compositae) is an herb primarily distributed throughout in the world. The species is regarded as ecologically important in the world. Safflower was used for medicines, as well as making red (carthamin) and yellow dyes. We have used the RAPD technique to investigate the phylogenetic relationships and genetic diversity of C. tinctorius. We obtained 123 bands from all the 26 cultivars. The average number of bands was 9.5 per primer. The genetic diversity of safflower is found among cultivars and there is a high among-cultivar differentiation. The OPC18-01 band is the specific marker for Syria cultivar, whereas no products were detected in individuals from other country cultivars. We found seven phenetic bands for determining the specific marker of cultivars with SCAR markers. Though the number of individuals sampled for analysis was small and probably not fully representative of the total available diversity in C. tinctorius, this study demonstrates that the regions (Morocco, Syria, and Turkey) of the Mediterranean Sea were more variable than other regions with the exception of India. In this result, although only simple result of RAPD is difficult to assert the center of species diversity of C. tinctorius, the regions of the Mediterranean Sea may be the most probable candidate for the origin of safflower. India was also the candidate of the center or secondary center of species diversity of C. tinctorius. RAPD markers were effective in classifying cultivar levels of safflower.

Analysis of Genetic Relationship of Entomogenous Fungi in Korea by Morphological Characteristics and RAPD (한국에서 채집한 Entomogenous fungi의 형태와 RAPD에 의한 유연관계 분석)

  • Choi, In-Young;Choi, Joung-Sik;Yu, Young-Jin;Lee, Wang-Hyu
    • The Korean Journal of Mycology
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    • v.29 no.1
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    • pp.34-40
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    • 2001
  • RAPD test and the observation of morphological, cultural characteristics of fourteen selected entomogenous fungi were conducted to investigate the analysis of their internal relationships. Paecilomyces tenuipes showed snow flower form attached to numerous white conidiophores, produced globular and semi-egg types on the club types of phialides. Cordyceps militaris formed globosely conidiophores, dark yellow fruiting body on pupa. The phialide as on Acremonium-type in global conidiophores. Beauveria bassiana covered with conidia was not formed fruiting body and adhered conidia on conidiophore of zigzag type. The PDA and SDAY medium were confirmed as an optimum growth of them. P. tenuipes showed to velvet and plane types in several media whereas C. militaris was belong to centrally raised and floccose in the morphological type. In contrast, B. bassiana covered with conidia on velvet shape. The size of amplified products were analyzed by RAPD using URP primer and were from 100 bp to 2.0 kb with $10{\sim}14$ geuomic DNA. Total similarities of two groups were by dendrogram of UPGMA analysis. The homology of P. tenuipes groups was 94.8 to 100%. It also showed 70.1 to 96.6% in C. militaris group and B. bassiana was higher similarity than any other. The internal change of C. militaris, produced telemorph fruiting body, was higher seperated in species than P. tenuipes and B. bassiana in the 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 Characteristics and Genetic Diversity Using the RAPD Technique in the Arksllell, Scapharca broughtosnii (Schrenck) Sronn Korea and China (한국산 및 중국산 피조개, Scapharca broughtonii(Schrenck)의 형태적 특성과 RAPD 기법을 이용한 유전적 분석)

  • LEE Jeong Mee;PARK Ji-Won;YOO Myung Suk;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.2
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    • pp.297-304
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    • 1997
  • Morphological and Benetic comparision were studied for the arkshell populations of Korea and China. The shell was collected from commercial arkshell bed in Jinhae Bay, Korea and from an hatchery in Young Seong, China. Shell Parameters, number of radial ribs and random amplified polymorphic DNA (RAPD) band were measured. The two populations had morphological and genetic differences. There was significant morphological difference $(P\leq0.05)$ in the ratio of the longest radial rib to the shell length. Posterior parts of the Chinese arkshell were more elongated than those of the Korean arkshell. Number of radial ribs were $36\~41$ for the Korean arkshell and $39\~43$ for the Chinese arkshell. RAPD markers generated tty each of 19 primers were $2\~6$ bands. Genetic diversity between the two Populations was clear since the genetic similarity was very low, not exceeding 0.29. The genetic similarity among the Korean arkshells $(0.58\~0.40)$ was higher than that of the Chinese arkshells $(0.48\~0.32)$.

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RAPD-SCAR Markers Linked to Medium-Leaf Zoysiagrass Ecotypes (한국잔디 중지 변이개체와 연관된 RAPD-SCAR 마커)

  • Chung, Sung Jin;Park, Su Jeong;Kim, Hun Joong;Yang, Geun-Mo;Choi, Joon-Soo;Oh, Chan-Jin;Jang, Deok-Hwan;Song, In-Ja;Lee, Geung-Joo
    • Weed & Turfgrass Science
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    • v.2 no.2
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    • pp.191-197
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    • 2013
  • Two medium-leaf ecotypes (CY6069, CY6097) belonging to one species (Zoysia japonica) of Korean lawngrasses were selected in sod production fields in Jang Seong, Korea. They were reported to have distinct morphological and growth rate characteristics different from the preferred medium-leaf type zoysiagrass in Korea. This study was conducted to define further the genotypic difference at the molecular level and to develop DNA marker based on the specific DNA fragment. Polymorphic DNA fragments were first explored by using randomly amplified polymorphic DNA (RAPD) primers, which were then converted into PCR-based sequence characterized amplified region (SCAR) markers. The CY6069-specific primer set amplified about 550 bp successfully, while the CY6097 marker produced the expected 690 bp band, by which those markers were nominated by CY6069_550 and CY6069_690 SCARs, respectively. Together with the reported morphological and other phenotypic features, the SCAR markers confirmed in this study will be useful to identify those medium-leaf zoysiagrass genotypes when they are cultivated with other vegetatively propagated warm-season turfgrasses in sod farms.

A taxonomic study of Abeliophyllum Nakai (O1eaceae) based on RAPD analysis (RAPD분석에 의한 미선나무속의 분류학적 연구)

  • 김동갑;박경량;김주환
    • Korean Journal of Plant Resources
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    • v.15 no.1
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    • pp.26-35
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    • 2002
  • Abeliophyllum distichum Nakai (O1eaceae) is a monotype of Korea and is distinguished from related genus Forsythia Vahl by the morphological characters such as fruit shape, flower color and etc. Even though several intraspecific taxa were reported according to the color of flowers and shape of fruits, there have been many controversals on the taxonomic indentity and status of rank on those taxa. In the present study, we performed the RAPD analysis to delimit the infraspecific taxa of Abeliophyllum distichum and to investigate the genetic polymorphism and relationships among 12 populations. 212 scorable RAPD markers with 70 common markers were found from the PCR reactions with 16 random oligoprimers and were analyzed by Nei's genetic distance. From 0.108 to 0.321 of genetic variations were showed among the taxa. Some regional groups instead of same taxa were clustered from the phonogram of UPGMA analysis. Also, we could not find distinct lineage among intraspecific taxa. The result from RAPD analysis supported that the infraspecific taxa of Abeliophyllum distichum might be the individual variations and treated as the same taxa. RAPD analysis was very useful to confirm the high gene pool with diverse genetic polymorphism among Abeliophyllum distichum populations.