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

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Development of strain-specific SCAR marker for selection of Pleurotus eryngii strains with higher β-glucan (베타글루칸 함량이 높은 큰느타리버섯 선발을 위한 SCAR marker 개발)

  • Kim, Su Cheol;Kim, Hye Soo;Cho, Yong Un;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.13 no.1
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    • pp.79-83
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    • 2015
  • In this study, SCAR marker that differentiates Pleurotus eryngii strains with higher ${\beta}$-glucan from control strain was developed. Genomic DNAs of 9 control strains of Pleurotus eryngii and 9 Pleurotus eryngii strains with higher ${\beta}$-glucan were analyzed by bulked segregant analysis (BSA) using randomly amplified polymorphic DNA (RAPD). One-hundred twenty RAPD primers were screened on bulked DNA samples and a unique DNA fragment with the size of 91 bp was yielded by OP-R03 primer from the Pleurotus eryngii strains with higher ${\beta}$-glucan. A sequence characterized amplified region (SCAR) marker, designated as OP-R03-1-F and OP-R03-1-R, was designed on the basis of the determined sequence. The PCR analysis with the OP-R03-1 primer showed that this SCAR marker can clearly distinguish the Pleurotus eryngii strains with higher ${\beta}$-glucan from the control strains.

Development of strain-specific SCAR marker for selection of Pleurotus eryngii strains adaptable to high-temperature (큰느타리버섯의 고온적응성 형질에 관련된 SCAR Marker 개발)

  • Kim, Su Cheol;Kim, Hye Soo;Park, So Yeon;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.226-231
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    • 2014
  • In this study, SCAR marker that differentiates Pleurotus eryngii strains adaptable to high-temperature from control strain was developed. Genomic DNAs of 7 control strains of Pleurotus eryngii and 7 Pleurotus eryngii strains adaptable to high-temperature were analyzed by bulked segregant analysis (BSA) using randomly amplified polymorphic DNA (RAPD). Onehundred twenty RAPD primers were screened on bulked DNA samples and a unique DNA fragment with the size of 385 bp was yielded by OP-A06 primer from the Pleurotus eryngii strains adaptable to high-temperature. A sequence characterized amplified region (SCAR) marker, designated as OP-A06-1-F and OP-A06-1-R, was designed on the basis of the determined sequence. The PCR analysis with the OP-A06-1 primer showed that this SCAR marker can clearly distinguish the Pleurotus eryngii strains adaptable to high-temperature from the control strains.

Geographic Variation in Pond Smelt (Hypomesus nipponensis) by RAPD Analysis (RAPD 분석에 의한 빙어 (Hypomesus nipponensis)의 지리적 변이)

  • Kim, Yong-Ho;Park, Su-Young;Yoon, Jong-Man
    • Korean Journal of Ichthyology
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    • v.18 no.1
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    • pp.1-11
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    • 2006
  • Genomic DNA isolated from two geographical populations of pond-smelt (Hypomesus nipponensis) was amplified for RAPD (randomly amplified polymorphic DNA) analysis. The populations were obtained from Chungju (CJ), in the inland area, and Dangjin (DJ), in the vicinity of the West Sea in Korea. Seven arbitrarily selected primers, OPB-06, OPB-10, OPB-13, OPB-17, OPC-09, OPC-17 and OPC-20, were used to generate the shared loci, polymorphic, and specific loci. Three hundred and eighty-three loci observed per primer were identified in the CJ population, and 287 were identified in the DJ population. Among them, 91 polymorphic loci or 23.8% were polymorphic in the CJ population, and 47 (16.4%) in the DJ population. The number of shared loci observed was 198 in the CJ population and 176 in the DJ population. Forty-four and 75 specific loci were detected in the CJ and DJ populations, respectively. Especially, 99 numbers of shared loci by the two populations, with an average of 14.1 per primer, were observed in the two pond-smelt populations. The average bandsharing value between the two geographical pond-smelt populations was $0.700{\pm}0.008$, ranging from 0.600 to 0.846. Compared separately, the bandsharing value of individuals within the CJ population was higher than that of the DJ population. The dendrogram obtained using the data from the seven primers indicated three genetic clusters: cluster 1, CJ 01, 02, 03, 04, 05, 06, 07, 08, 09, 10, and 11; cluster 2, DJ 01, 02, 03, 04, 05, 06, 07, 08, and 09; and cluster 3, DJ 10 and 11. The genetic distance between the two geographical populations ranged from 0.040 to 0.545. Thus, RAPD-PCR analysis revealed a significant genetic distance between the two pond-smelt populations.

Induction of Somatic Embryos and RAPD Analysis in Regenerated Plantlets of Bupleurum falcatum L. (자호(紫胡)의 체세포배(體細胞胚) 형성(形成)과 재생(再生) 식물체(植物體)의 RAPD 분석(分析))

  • Park, Cheol-Ho;Yu, Chang-Yeon;Seo, Jeong-Sik;Kim, Ki-Sik;Ahn, Sang-Duek;Chang, Byoung-Ho
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.1
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    • pp.50-55
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    • 1995
  • This study was conducted to determine the optimum conditions for induction and different somatic of somatic embryos as well as germination of encapsulated and stored somatic embryos. Somatic embryos was better formed in 1/2X MS medium than full - strength MS medium. 0.1 to 1.0mg/lBA and kinetin promoted shoot differentiation of somatic embryos. Higher concentration tend to inhibit differentiation. IAA affect positively both root and shoot growth. In vitro germination of somatic embryos encapsulated with 2% alginate matrix containing 1/2 MS nutrient medium and $AgNO_3$ 5mg/l was 86%. Storage of somatic embryos was effecive at $5^{\circ}C$ but the germination rate decreased with longer storage period. RAPD analyses with plants regenerated from the somatic embryos showed DNA polymorphism, indicating abolition of primer binding site by point mutation, deletion, or insertion of certain sequences.

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Physiological Properties of Two Japonica Rice (Oryza sativa L.) Cultivars: Odae and Ilpum

  • Cho, Hye-Jeong;Heo, Kweon;Umemoto, Takayuki;Wang, Myeong-Hyeon
    • Journal of Applied Biological Chemistry
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    • v.50 no.3
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    • pp.127-131
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    • 2007
  • The properties of two cultivars of japonica rice, Odae (early ripening variety) and Ilpum (late ripening variety), were compared. They grew on MS (Murashige and Skoog) medium but the growth of both cultivars was strongly retarded by 50 mM or more salt. There was no clear difference between the growths of seedlings of the two cultivars for the first 24 h after germination. The amylopectin chain-length profiles of the two cultivars did not differ significantly, and amylopectin content was estimated at $16.0{\pm}0.4%$ in cv. Odae and $16.4{\pm}0.4%$ in cv. Ilpum. A total of 114 RAPD (randomly amplified polymorphic DNA) fragments ranging from 0.4 to 2.5 kb were isolated from the two cultivars, 61 from cv. Odae and 53 from cv. Ilpum, indicating that there is little genetic variation between them.

Genotypic Characterization of Oak Wilt Pathogen Raffaelea quercus-mongolicae and R. quercivora Strains (참나무류에 시들음병을 일으키는 Raffaelea quercus-mongolicae와 R. quercivora의 유전적 특성)

  • Seo, Sang-Tae;Kim, Kyung-Hee;Lee, Sang-Hyun;Kwon, Yong-Nam;Shin, Chang-Hoon;Kim, Hye-Jeong;Lee, Sang-Yong
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.219-223
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    • 2010
  • Recently, the oak wilt diseases especially on Quercus mongolica, have been increasing in various districts of Korea. A collection of 38 strains of the oak wilt pathogen Raffaelea qeurcus-mongolicae and R. quecivora isolated from Quercus spp. in Korea and Japan was characterized by $\beta$-tubulin gene sequence and randomly amplified polymorphic DNA (RAPD) analysis. In cluster analysis based on $\beta$-tubulin gene sequence the strains were divided into 4 clusters, of which clusters 2 and 4 were composed of Japanese strains except for one Korean strain. RAPD analysis showed that they were also effectively differentiated by a strong RAPD fragments. On the basis of the two genetic analysis, significant differences were detected between Korean strains and Japanese strains.

Genetic diversity and herbicide resistance of 15 Echinochloa crus-galli populations to quinclorac in Mekong Delta of Vietnam and Arkansas of United States

  • Le, Duy;Nguyen, Chon M.;Mann, Richard K.;Yerkes, Carla N.;Kumar, Bobba V.N.
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.472-477
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    • 2017
  • Barnyardgrass (Echinochloa crus-galli) is one of the worst weeds in rice (Oryza sativa), but there are few reports about the genetic diversity and herbicide resistance of barnyardgrass in Vietnam. In this study, we used random amplified polymorphic DNA (RAPD) analysis and greenhouse testing to study the genetic diversity and quinclorac resistance levels of 15 Echinochloa crus-galli populations in the Mekong Delta, Vietnam, and the state of Arkansas, U.S. The quinclorac resistance of Echinochloa crus-galli populations in Vietnam was confirmed; 9 populations were resistant to quinclorac with R/S ratios ranging from 1.9 to 6.3. Six oligonucleotide primers produced a total of 55 repeatable bands of which 46 were polymorphic (83.3% average) among the 15 populations. Genetic distance was calculated, and cluster analysis separated the 15 populations into 2 main clusters with the genetic distances within the clusters ranging from 0.09 to 0.39. The two main clusters were divided into 7 subclusters, and the quinclorac resistant and susceptible populations were located randomly within each subcluster. Six out of 13 weed populations from Vietnam belonged to one cluster and a single Echinochloa species. The remaining 7 populations were identified as potentially different species in the Echinochloa genus. Nine Echinochloa populations from Vietnam were tested and identified as quinclorac resistant. The connection between quinclorac resistance levels and weed groups defined by RAPD analysis in the study is unclear; the quinclorac resistance of each resistant population could have evolved individually, regardless of differences in genetic diversity and location of the sampled populations.

Development of a psychrophilic-SCAR marker for Pleurotus eryngii (큰느타리버섯의 저온적응성 형질에 관련된 SCAR Marker 개발)

  • Kim, Su Chul;Hwang, Hye Sung;Cho, Yun Jun;Kim, Hye Su;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.11 no.3
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    • pp.171-176
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    • 2013
  • Genomic DNAs of psychrophilic strains of Pleurotus eryngii were analyzed by randomly amplified polymorphic DNA (RAPD) using OP-A, OP-B, OP-L, OP-P, OP-R and OP-S3 primers to develop the strain-specific DNA marker. A unique DNA fragment with the size of 480 bp was yielded by OP-S3 primer from the psychrophilic strain. A sequence characterized amplified region (SCAR) marker, designated as OP-S3-1, was designed on the basis of the determined sequence. The PCR analysis with the OP-S3-1 primer showed that this SCAR marker can clearly distinguish the psychrophilic strains from the control strains.

RAPD Analysis for the Evaluation of Genetic Diversity Among the Fusarium Species from Various Sources (각종 작물로부터 분리한 Fusarium속 균의 RAPD 기법을 이용한 유전분석)

  • Choi, Hei-Sun;Kim, Kyong-Soo;Kim, Myong-Jo;Shim, Jae-Ouk;Kim, Byong-Sup;Lee, Min-Woong;Lee, Youn-Su
    • The Korean Journal of Mycology
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    • v.25 no.3 s.82
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    • pp.202-208
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    • 1997
  • In this study, we evaluated the use of RAPD method to discriminate among strains of Fusarium species including F. oxysporum and f. sp. of F. oxysporum. As a result of the amplication, fifteen primers showed total 180 bands ranging from 0.2 to 3 Kb. Among those 180 bands, 126 polymorphic bands were used for bionominal matrix code (0, 1), and UPGMA dendrogram analysis. Fusarium oxysporum isolate 355 showed high similarity with F. oxysporum isolate 358 at 0.9603. Fusarium roseum isolate 87 and F. oxysporum isolate 358, F. o. f. sp. lycopersici isolate 69 and F. o. f. sp. melongena 68 showed low similarity of 0.3809. Fusarium oxysporum isolate 361 and F. o. f. sp. raphani isolate 218 showed similarity of 0.8730, F. oxysoprum isolate 354 and unidentified Fusarium sp. isolate 228 showed similarity matrix of 0.7936, and F. roseum isolate 87 and F. o. f. sp. raphani isolate 57 showed similarity matrix of 0.5873.

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Selection of PCR Markers and Its Application for Distinguishing Dried Root of Three Species of Angelica

  • Jin, Dong-Chun;Sung, Jung-Sook;Bang, Kyong-Hwan;In, Dong-Su;Kim, Dong-Hwi;Park, Hee-Woon;Seong, Nak-Sul
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.2
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    • pp.121-125
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    • 2005
  • An analysis of RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) was performed with three Angelica species (A. gigas Nakai, A. sinensis (Olive.) Diels and A. acutiloba Kitag) in an effort to distinguish between members of these three species. Two arbitrary primers (OPC02, OPD11) out of80 primers tested, produced 17 species-specific fragments among the three species. Eight fragments were specific for A. sinensis, four fragments specific for A. gigas, five specific for A. acutiloba. When primers OPC02 and OPD11 were used in the polymerase chain reaction, RAPD-PCR fragments that were specific for each of the three species were generated simultaneously. Primer OPC02 produced eight species-specific fragments: four were specific for A. sinensis, one for A. gigas, and three for A. acutiloba. Primer OPD11 produced nine speciesspecific fragments: four for A. sinensis, three for A. gigas, and two for A. acutiloba. The RAPD-PCR markers that were generated with these two primers should rapidly identify members of the three Angelica species. The consistency of the identifications made with these species-specific RAPD-PCR markers was demonstrated by the observation that each respective marker was generated from three accessions of each species, all with different origins. We also performed the RAPD-PCR analysis with the dried Angelica root samples that randomly collected from marketed and from the OPC02 primer, obtained a A. gigasspecific band and the band were cloned and sequenced.