• Title/Summary/Keyword: RAPD PCR

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Random Amplified Polymorphic DNA (RAPD) Variation in Porhyra yezoensis and P. tenera

  • Beom-Kyu Kim;Gyu-Hwa Chung;Yuji Fujita
    • Journal of Aquaculture
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    • v.10 no.3
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    • pp.321-326
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    • 1997
  • The random amplified polymorphic DNA (RAPD) technique was used to anlayze six isolates of two species of Porphyra, P. yezoensis and P. tenera. Four 21-mer prrmers were combined randomly into six groups of double primers and screened for DNA amplification using nuclear and chloroplast tempate DNA. The RAPD patterns resulting from RnRc and CnCc primers provided evidence for both genetically homo-and heterogeneous populations of P. yezoensis and P. tenera. Similarity values obtained by RnRc primer analysis of nuclear DNA varied from 0.364 to 0.714 and those of chloroplast DNA were high, ranging from 0.727 to 1.000, except for P. yezoensis (Enoura).

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Genetic Similarity and Difference between Common Carp and Israeli Carp (Cyprinus carpio) Based on Random Amplified Polymorphic DNAs Analyses

  • Yoon, Jong-Man
    • Animal cells and systems
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    • v.5 no.4
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    • pp.333-339
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    • 2001
  • Common carp (Cyprinus carpio) and its aquaculture breed Israeli carp samples were obtained from two separate aquaculture facilities under the similar raising conditions during two years in the Kunsan National University, Korea. Genomic DNA was isolated from the common carp and Israeli carp for identification of genetic characteristics and genomic polymorphisms by polymerase chain reaction amplification of DNA using arbitrary primers. The arbitrary primer No.21 (ACTTCGCCAC) yielded the highest number of fragments with the average of 15.0 among the primers used in Israeli carp. A tota1 of 294 polymorphic products in common carp and 336 in Israeli carp were observed by random primers. The average number of polymorphic products generated by random RAPD primer No. 2 (GTAGAC-CCGT) showed 8.0 in Israeli carp. On average, each random RAPD primer produced 5.4 amplified polymorphic products in common carp and 6.2 in Israeli carp. An average genetic similarity (BS value) was 0.44$\pm$0.05 within the common carp and 0.32$\pm$0.04 within the Israeli carp. The degree of similarity frequency (BS) between two carps was 0.67 as generated by the primer No. 19 (GACGGATCAG). The average level of bandsharing was 0.57$\pm$0.03 between the two carps. Accordingly, the two carp populations were genetically a little distant. The electrophoretic analysis of PCR-RAPD products showed middle levels of variation between the two carp populations. This result implies that the genetic diversity among intra-population may be higher when compared with that between the two carps. The RAPD polymorphism generated by these random primers might be used as a genetic marker for populations or lines identification in important aquacultural carp.

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Genetic characterization of microsporidians infecting Indian non-mulberry silkworms (Antheraea assamensis and Samia cynthia ricini) by using PCR based ISSR and RAPD markers assay

  • Hassan, Wazid;Nath, B. Surendra
    • International Journal of Industrial Entomology and Biomaterials
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    • v.30 no.1
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    • pp.6-16
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    • 2015
  • This study established the genetic characterisation of 10 microsporidian isolates infecting non-mulberry silkworms (Antheraea assamensis and Samia cynthia ricini) collected from biogeographical forest locations in the State of Assam, India, using PCR-based markers assays: inter simple sequence repeat (ISSR) and random amplified polymorphic DNA (RAPD). A Nosema type species (NIK-1s_mys) was used as control for comparison. The shape of mature microsporidian spores were observed oval to elongated, measuring 3.80 to $4.90{\mu}m$ in length and 2.60 to $3.05{\mu}m$ in width. Fourteen ISSR primers generated reproducible profiles and yielded 178 fragments, of which 175 were polymorphic (98%), while 16 RAPD primers generated reproducible profiles with 198 amplified fragments displaying 95% of polymorphism. Estimation of genetic distance coefficients based on dice coefficients method and clustering with un-weighted pair group method using arithmetic average (UPGMA) analysis was done to unravel the genetic diversity of microsporidians infecting Indian muga and eri silkworm. The similarity coefficients varied from 0.385 to 0.941 in ISSR and 0.083 to 0.938 in RAPD data. UPGMA analysis generated dendrograms with two microsporidian groups, which appear to be different from each other. Based on Euclidean distance matrix method, 2-dimensional distribution also revealed considerable variability among different identified microsporidians. Clustering of these microsporidian isolates was in accordance with their host and biogeographic origin. Both techniques represent a useful and efficient tool for taxonomical grouping as well as for phylogenetic classification of different microsporidians in general and genotyping of these pathogens in particular.

Analysis Genetic Similarity of Gentiana scabra var. buergeri by Randomly Amplified Polymorphic DNA (RAPD를 이용한 용담의 유전적 유사도 분석)

  • Lee, Hae-Kyung;Lee, Mi-Kyung;Moon, Chang-Sik;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.4 no.3
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    • pp.224-230
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    • 1996
  • Randomly amplified polymorphic DNA(RAPD) analysis was applied to detect the molecular polymorphisms in Gentiana scabra var. buergeri. A high level of molecular variability was found among wild plants and cultivars. In genetic analysis, eight of twenty primers were selected and 54 amplification products ranged 2. 2 to 0.2 kb were compared. Twenty - nine amplified products showed polymorphic, while five were monomorphic. Twenty of line specific bands were found. In genetic similarity and cluster analysis using PCR products, three wild plants collected from Naejangsan, Daedunsan and Keojedo and one cultivar Seochunjaerae were grouped into one cluster, while cultivar Jinbujaerae and Japanese line separated into another clusters, respectively. The identification of DNA polymorphisms by the RAPD technique will facilitate the selection of the lines from different origin.

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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.

Analysis of phylogenetic relationship among Korean Pinellia Tenore (Araceae) using RAPD markers (RAPD markers에 의한 한국산 반하속 식물의 유연관계 분석)

  • Tae, Kyoung-Hwan;Kim, Dong-Kap;Kim, Joo-Hwan
    • Korean Journal of Plant Taxonomy
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    • v.35 no.3
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    • pp.161-174
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    • 2005
  • In order to presume the relationships between two species of P. ternata and P. tripartia, and their populations of the Korean Pinellia, RAPD analysis was performed. The length of the amplified DNA fragments ranged from 300 to 2,500bp. Seventy scorable RAPD makers were found from the PCR reactions with 7 random oligoprimers and were analyzed by Nei-Li's genetic coefficient. Also, some regional groups instead of same taxa were clustered from the phenogram of UPGMA analysis and NJ tree. Populations within each species were clustered at low genetic distance, there had the closed relationship. According to the regional individuals, Pinellia ternata was showed the variation pattern of morphological (leaf shape and flower color) and cytological characters(somatic chromosome numbers). So we suggested to difference of characteristic variety based on variety of habitat. According to this study, new species (Pinellia sp.) was affiliated with Pinellia and had the closest relationship with Hallasan and Japan population. The RAPD data was very useful to define the genetic variation and to discuss the relationships among the intraspecific taxa and their populations of the Korean Pinellia.

Genetic Relationship among Sedum Species Based on Morphological Characteristics and RAPD Analysis (한국산 Sedum속 식물의 형태적 특성과 RAPD에 의한 유연관계 분석)

  • Kwon, Soon Tae;Jeong, Jeong Hag
    • Horticultural Science & Technology
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    • v.17 no.4
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    • pp.489-493
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    • 1999
  • This study was conducted to investigate the potentiality of various Korean Sedum species as ornamental plants based on morphological characteristics and to analyze the genetic relationship among the Sedum species. S. kamtschaticum and S. takesimense possessing splendour flowercluster with yellow color could be suggested for garden plant, S. routundifolium having pink flower-clusters with round leaf shape for pot flower or garden plant and S. sarmentosum, S. polystichoides and S. oryzifolium with creeping stem and low plant height for ground cover plant or floral carpet. Eighteen oligonucleotide random primers were used to amplify genomic DNA of Sedum species using polymerase chain reaction (PCR). Ninety five polymorphic bands among 125 different DNA fragments in the range of 224 to 3,675 base pairs were obtained from RAPD analysis. Similarity matrix of RAPD profiles was generated by coefficient value of variation, and the data were subjected to be cluster analysis. Fifteen lines of Sedum species analyzed were classified into 3 groups with the similarity coefficient value of 0.418, and 12 groups with the value of 0.328. RAPD results showed similar trends as the morphological characteristics of the plants.

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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.

Pholiota adiposa and its Related Species Collected from the Wild Forestry (야생에서 채집된 검은비늘버섯(Pholiota adiposa)균에 관한 연구)

  • Lee, Sang-Sun;Kim, Mi-Hye;Chang, Hu-Bong;Shin, Chun-Sik;Lee, Min-Woong
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.574-582
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    • 1998
  • Five basidiocarps of Pholiota species have been collected from the areas of BubJu Temple for last two years, and identified to those of P. adiposa or Pholiota species. The taxonomy of these basidiocarps with the morphological aspects was compared with the analysis obtained from the polymorphisms of PCR-RAPD bands made after reacted with the random primers. The polymorphic variations were observed within the species of the basidiocarps, but not between genomic DNA's of the mycelia obtained and the basidiocarps. Several different bands made from the primers (28 and 36) in PCR-RAPD reactions were identified within the genus of Pholiota and speculated to be specific for the individual basidiocarp of P. adiposa collected. The primers employed here were considered to be very useful for distinguishing the individual isolates or basidiocarps collected from the fields. Also, the basidiospores were obtained from the sporeprints of the above basidiocarps as a simple agar and confirmed with observations of clamp connection under microscopes. The matings of them indicated the 'tetrapolar' type, being different from the 'bipolar' type reported by Japanese basidiocarps of P. adiposa or P. nameko. Based on our work, the edible fungi collected were speculated to be a new breeding resource for those of Pholiota commercialized in Japan.

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Intraspecific Variations of Rubus oldhami (덩굴딸기(Rubus oldhami)의 종내변이)

  • Kim Mi-Na;Jang Su-Kil;Yoo Ki-Oug
    • Korean Journal of Plant Resources
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    • v.18 no.2
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    • pp.260-269
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    • 2005
  • Taxonomic studies on morphological, principal component analysis (PCA), palynological, RAPD and PCR-RELP analysis were conducted to intraspecific relationships of Rubus oldhami. Three types of Rubus oldhami based on the flower characters such as petal length and number were used in this study. Among the 14 morphological characters, perianth length, calyx lobe length, apical leaflet shape and leaflet length were used to distinguish for each type. The pollen characters such as shape, aperture number, surface sculpture were showed very similar among three types. Eight primers out of 20 arbitrary primers were screened for three types, and were revealed 33 ($60\%$) polymorphic bands. The phonogram by RAPD data showed incongruent with morphological analysis. Even though ten restriction endonucleases produced 20 restriction sites, polymorphic bands were not observed. Based on the results, three types of Rubus oldhami divided well by morphological characters, but pollen and DNA data were not supported. Therefore, type 1 and 2 which different from type 3 by flower characters considered as a temporary hybrid or ecotype because of their similar habitats.