• Title/Summary/Keyword: RAPD primer analysis

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Genetic Variability Based on Randomly Amplified Polymorphic DNA in Mistletoe Fig (Ficus deltoidea Jack) Collected from Peninsular Malaysia

  • Bhore, Subhash Janardhan;Arneida H., Nurul;Shah, Farida Habib
    • Journal of Forest and Environmental Science
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    • v.25 no.1
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    • pp.57-65
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    • 2009
  • Ficus deltoidea Jack is an important and popular medicinal plant species found in the Malaysia. Plants are being collected and used based on morphology and authentication to prevent adulteration is not in practice. In this study, twenty-six accessions of F. deltoidea Jack were collected from Kelantan and Terengganu states of Peninsular Malaysia to examine their genetic similarities and differences using randomly amplified polymorphic DNA (RAPD) technique. Out of 20 arbitrary primers, two primers (D-10 and D-11) were selected which produced reliable DNA polymorphism. D-10 and D-11 primers generated 138 RAPD bands ranging from 250 bp to 3000 bp. Ninety-nine of them were polymorphic loci (72%) and thirty-nine were nonpolymorphic loci (28%). A total of 56 bands with polymorphic loci were amplified with primer D-10 and analyzed by cluster analysis and UPGMA to present a dendrogram depicting the degree of genetic relationship among 26 accessions. Eight RAPD markers were sequenced to determine their identity. RAPD analysis showed the genetic diversity among 26 accessions of F. deltoidea Jack. The RAPD profile and RAPD marker sequences reported in this paper could be used in plant and/or plant material authentication. This study also suggested that RAPD can be a useful technique to study DNA polymorphism in F. deltoidea Jack.

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Parameters Affecting Polymerase Chain Reaction in RAPD Analysis of Pleurotus spp. (느타리버섯속(屬)의 DNA 다형성분석(多型性分析)에 영향(影響)을 미치는 PCR 조건(條件))

  • Kim, Beom-Gi;Jeong, Mi-Jeong;Lee, Chang-Soo;Lee, Hee-Kyung;Yoo, Young-Bok;Ryu, Jin-Chang
    • The Korean Journal of Mycology
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    • v.23 no.3 s.74
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    • pp.202-208
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    • 1995
  • This study describes the effects of several components on PCR amplification used for RAPD. We used different concentrations of reaction components to obtaine discrete and reproducible PCR products from Pleurotus cornucopiae. The optimum concentrations of reaction components were found to be 80 ng of template DNA, 30 pmole of 10-mer primer, $200\;{\mu}M$ dNTP, 2mM $MgCl_2$, 50 mM KCl, 10 mM Tris-HCl(pH 9.0), 0.1% Triton X-100, 1.5 unit of Taq DNA polymerase (promega) in $50\;{\mu}l$ reaction volume. The optimum annealing temperature was $35^{\circ}C$. These results proved to be valuable for characterization of Pleurotus spp.

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Genetic Variation in Fusarium oxysporum f. sp. fagariae Populations Based RAPD and rDNA RFLP Analyses

  • Nagaraian, Gopal;Nam, Myeong-Hyeon;Song, Jeong-Young;Yoo, Sung-Joon;Kim, Hong-Gi
    • The Plant Pathology Journal
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    • v.20 no.4
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    • pp.264-270
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    • 2004
  • Fusarium oxysporum f. sp. fragariae is a fungal pathogen causing strawberry wilt disease. The random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphisms (RFLPs) of intergenic spacer (IGS) region of rDNA were used to identify genetic variation among 22 F. oxysporum f. sp. fragariae isolates. All isolates could be distinguished from each other by RAPD analysis and RFLP of 2.6 kb amplified with primer CNS1 and CNL12 for IGS region of rDNA. Cluster analysis using UPGMA showed eight distinct clusters based on the banding patterns obtained from RAPD and rDNA RFLP. These results indicate that F. oxysporum f. sp. fragariae isolates are genetically distinct from each other, There was a high level genetic variation among F. oxysporum f. sp. fragariae.

Assessment of Genetic Relationship among Curcurbitaceae Cultivars Revealed by RAPD Marker (RAPD Marker에 의한 호박의 품종간 유연 관계 분석)

  • 김창훈;이승인;유병천;송인호;권용삼
    • Journal of Life Science
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    • v.13 no.5
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    • pp.590-595
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    • 2003
  • The objective of this study was to assess of genetic variation within and between pumpkin species including Cucurbita maxima, C. moschata, C. pepo and C. maxima${\times}$C. moschata using RAPD markers. The 16 primers showed the amplification of 136 scorable fragments ranging from about 100 bp to 2300 bp. A total of 94 DNA fragments were polymorphic with an average 5.9 polymorphic bands per primer. A species $(C. maxima\timesC. moschata)$ has the highest number of polymorphic loci. Based on obtained data, UPGMA cluster analysis was conducted. Twenty pumpkin cultivars were classified into three large categories and identified genetic distance of cluster ranging from 0.38 and 1.00. Clustering was in accordance with the division of Curcurbitaceae into four species, C. maxima, C. moschata, C. pepo and C. $C. maxima\timesC. moschata$. Therefore, RAPD method may be essential tool for enabling discrimination of pumpkin cultivars.

Genetic Polymorphisms and phylogenetic Relationships of Italian Ryegrass Cultivars Based on Random Amplified Polymorphic DNA ( RAPD ) Markers (RAPD 표지인자를 이용한 이탈리안 라이그라스 품종의 유전적 변이 및 유연관계 분석)

  • 임용우;이승재;신정섭;정영수;최기준;임영철;임근발;박병훈
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.18 no.1
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    • pp.35-42
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    • 1998
  • Eleven Italian ryegrass cultivars were examined for their genetic polymorphisms and phylogenetic relationships using randomly amplified polymorphic DNA (RAPD) markers. In RAPD analysis of 34 random primers, 96 of total 162 bands obtained from 16 primers were polymorphic and sizes of polymorphic band ranged between 0.5 and 1.5kb. Number of bands amplified per primer was varied from 3 to 16 and average number was 14.8. Phylogenetic relationship among cultivars based on the RAPD analysis was examined using UPGMA computer program. In pairwise genetic similarity test of 11 Italian ryegrass cultivars, Grazer and Orlando showed highest coefficient of genetic similarity as 0.740, whereas Marshall and Orlando was lowest as 0.438. Eleven Italian ryegrass cultivars were grouped into 3 major clusters and genetic distance of clusters ranged between 0.567 and 0.646, indicating low level of genetic variation.

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Differentiation of Lentinus edodes Isolates in Korea by Isozyme Polymorphisms and Random Amplified Polymorphic DNA (RAPD) Analysis (Isozyme Polymorphism 및 Random Amplified Polymorphic DNA(RAPD) Pattern에 의한 표고 버섯 품종간 비교)

  • Park, Won-Mok;Ko, Han-Gyu;Park, Ro-Jo;Hong, Ki-Sung;Kim, Gyu-Hyun
    • The Korean Journal of Mycology
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    • v.25 no.3 s.82
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    • pp.176-190
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    • 1997
  • Sixty-three isolates of Lentinus edodes obtained from Korea were used to assess the genetic similarity by isozyme polymorphisms and random amplified polymorphic DNA patterns. The activities of esterase, peroxidase and acid phosphatase displayed 10, 7 and 3 distinct isozyme patterns, respectively. By combining the isozyme patterns obtained with the 3 enzymes, every isolate showed its own distinct electrophoretic phenotypes. A distance matrix calculated between all pairs of 63 electrophoretic phenotypes based on the presence or abscence of isozyme bands were analyzed by the group-average method. Results of the cluster analysis assinged the 63 phenotypes into six major groups. In the analysis of random amplified polymorphic DNA patterns, all isolates of Lentinus edodes were devided into five RAPD groups.

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Genetic Relationship among Garlic Cultivars Based on RAPD Analysis (RAPD에 의한 마늘의 유연관계 분석)

  • 권순태;오세명
    • Journal of Life Science
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    • v.9 no.6
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    • pp.671-676
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    • 1999
  • RAPD analysis using random primers were tried to evaluate the genetic variation and diversity of the nine garlic cultivars including two foreign varieties. Thirty-two primers out of 70 primers screened were used to amplify genomic DNA of garlic cultivars using polymerase chain reaction(PCR). Among a total of 151 bands amplified by 32 primers, 125 polymorphic bands were subjected to analysis for genetic relationship of garlic cultivars. The estimated size of amplified PCR products were in the range of 932 to 4,060 base pairs. Nine garlic cultivars were classified into two groups, such as group I corresponded to Changnyung and Hungary cultivars, and group II, Namdo, Sandong from China, Yecheon, Euiseong, Youngweol, Danyang, Jeongsun cultivars, with the genetic distance value of 0.271. The major ecological types of garlics, so called southern and northern types, was grouped in the genetic distance value of 0.200. The results presented in this study suggest that RAPD analysis are likely to be useful for identification of cultivars and evaluation of genetic origin in garlics.

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DNA Fingerprinting of Rice Cultivars using AFLP and RAPD Markers

  • Cho, Young-Chan;Shin, Young-Seop;Ahn, Sang-Nag;Gleen B. Gregorio;Kang, Kyong-Ho;Darshan Brar;Moon, Huhn-Pal
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.1
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    • pp.26-31
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    • 1999
  • This experiment was conducted to evaluate genetic variation in 48 rice accessions (Oryza sativa L.) using AFLP and RAPD markers. For AFLP, a total of 928 bands were generated with 11 primer combinations and 327 bands (35.2%) of them were polymorphic among 48 accessions. In RAPD analyses using 22 random primers 145 bands were produced, and 121 (83.4%) were polymorphic among 48 accessions. Each accession revealed a distinct fingerprint by two DNA marker systems. Cluster analysis using AFLP-based genetic similarity tended to classify rice cultivars into different groups corresponding to their varietal types and breeding pedigrees, but not using RAPD-based genetic similarity. The AFLP marker system was more sensitive than RAPD in fingerprinting of rice cultivars with narrow genetic diversity.

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Analysis of Diversity of Panax ginseng Collected in Korea by RAPD Technique (RAPD 방법을 이용한 국내 수집 인삼 (Panax ginseng C. A. Meyer)의 다양성 분석)

  • Seo, Sang-Deog;Yuk, Jin-Ah;Cha, Sun-Kyung;Kim, Hyun-Ho;Seong, Bong-Jae;Kim, Sun-Ick;Choi, Jae-Eul
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.5
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    • pp.377-384
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    • 2003
  • Genetic differences among nine land races of Korean ginseng (Panax ginseng C. A. Meyer) were examined using RAPD markers. Land races of Korean ginseng were collected from nine regions in Korea: Cheongwon, Guesan, Geumsan, Namwon, Pochun, Yangju, Yeoncheon, Yeongju. Out of 48 RAPD primers tested, 5 primers (OPA 7, OPA 13, URP 2, URP 3 and UBC 3) produced remarkable bands which showing polymorphisms among evaluated collections. Lower levels of genetic diversity were in detected same land races than among other land races. Genetic differences within and among land races indicate heterogeneity. These results indicate that cultivated ginseng in Korea is heterogeneous. Genetic similarity matrices of RAPD profiles were generated via coefficients of variation and the data were processed by the cluster analysis (UPGMA). When 90 collections were evaluated using selected 5 primers, those were clustered to 5 and 3 subgroups. These differences in genetic variation between land races of Korean ginseng implied the potential source for further breeding of Korean ginseng.

Genetic Diversity and DNA Polymorphism in Platycodon grandiflorum DC. Collected from East-Asian Area

  • Park, Chun-Geun;Yan, Zhi-Yi;Lee, Sang-Chul;Shon, Tae-Kwon;Park, Hee-Woon;Jin, Dong-Chun
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.2
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    • pp.115-120
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    • 2005
  • Broadening the genetic base of Platycodon grandiflorum DC. cultivar to sustain improvement requires assessment of genetic diversity available in P. grandiflorum DC.. The objective of this study was to analyze the genetic variation, genetic relationship among 48 samples collected from East-Asian Area by means of RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) markers. From the 18 primers tested, produced total 211 bands with an average of 11.7 bands per primer and obtained 103 polymorphic band with an average of 5.7 bands per primer,s revealed relatively high percentage of polymorphic bands (48.8%). The genetic similarities calculated from RAPD data varied from 0.688 to 0.994 and were clustered to six major groups on a criterion of 0.78 similarity coefficient. The present study has revealed the significant genetic similarity among the samples tested. The analysis of genetic relationships in P. grandiflorum using RAPD-PCR banding data can be useful for the breed improvement.