• Title/Summary/Keyword: RAPD primer

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Phylogenetic relationships in different strains of Pholiota species based on PCR polymorphism (PCR 다형성 분석에 의한 비늘버섯 속 계통의 유연관계 분석)

  • Kwon, Woon-Hyuk;Park, Hyuk;Baek, Min-Jae;Cho, Woo-Jin;Choi, Woo-Jeong;Ahn, Chi-Beom;Shin, Do-Bin;Lee, Tae-Soo
    • Journal of Mushroom
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    • v.11 no.2
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    • pp.69-76
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    • 2013
  • Pholiota species were collected from different geographical regions of the world. Genetic diversity and phylogenetic relationships were analyzed by rDNA-ITS sequences and RAPD polymorphism. The sizes of rDNA-ITS PCR amplicons of Pholiota spp. varied from 233~271, 158~223 and 174~219 bp, respectively. A phylogenetic tree was constructed on the ITS region sequences and Pholiota strains were classified into 8 clusters. Twenty strains in seven Pholiota spp. were classified into seven clusters by RAPD polymorphism using 15 arbitrary primers. Our experimental results suggested that rDN-ITS and RAPD analysis are useful tool for classifying Pholiota spp. and strains.

Genetic Variability of Pleurotus ostreatus Monospore Isolates by Random Amplified Polymorphic DNA Analysis (DNA 다형성 분석에 의한 느타리버섯 단포자 분리주의 유전적 변이성)

  • Song, Yeong-Jae;Jeong, Mi-Jeong;Kim, Beom-Gi;Rho, Yeong-Deok;Ryu, Jin-Chang;Yoo, Young-Bok
    • The Korean Journal of Mycology
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    • v.24 no.3 s.78
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    • pp.186-205
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    • 1996
  • This study was carried out to obtain data concerning the genetic variability of Pleurotus ostreatus. Monospores of P. ostreatus were isolated and cultured to estimate differences in the rate of mycelial growth and genetic similarity among the isolates. Although the overall growth rates were slow compared to their parental dikaryon except 2-MI 4, significant differences in the rate of mycelial growth were observed among isolates. Random amplified polymorphic DNA (RAPD) analysis using twenty six random primers showed 345 polymorphic DNA bands from 35 monospore isolates and their parental dikaryon. DNA bands ranged from 0.1 to 4.0 Kb in size. Most various polymorphic DNA bands within monospore isolates were obtained when we used J (OPA-01) or W (OPB-04). The 36-MI 103 showed totally different band patterns from those of the others. RAPD analysis revealed that there is approximatly 75% genetic similarity between monospore isolates and their parental dikaryon except 36-MI 103, which showed only 49% genetic similarity. In addition, genetic similarity degrees were classified into four groups: I (parental dikaryon), II (fast growing type), III (moderate growing type), and IV (slow growing type). However, there is no correlation between mating type and mycelial growth rates.

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Development of a Female-associated SCAR Marker in Schisandra nigra Max. (Schisandra nigra Max.에서 암그루에 연관된 SCAR 마커의 개발)

  • Han, Hyo Shim;Jung, Jae Sung
    • Journal of Life Science
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    • v.31 no.6
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    • pp.537-542
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    • 2021
  • Schisandra nigra Max., a dioecious plant native to Jeju Island in Korea, is cultivated on a small scale for fruit production. As fruit-producing female individuals are generally considered to be more valuable than male, early identification of plant sex at the seedling stage is important. In this study, a sequence-characterized amplified region (SCAR) marker associated with a female-specific region in the genome of S. nigra was investigated. Of 120 randomly amplified polymorphic DNA (RAPD) primers, one primer (OPB-03) consistently amplified a 749 bp band in female plants. The female-specific PCR product was isolated and cloned, and the nucleotide sequences were then determined. Southern hybridization performed using the female-specific fragment as a probe produced positive signals only in genomic DNA from the female plants. This result revealed that the 749 bp segment of DNA was present in the genome of female plants but absent in the genome of male plants. A SCAR primer pair was designed based on the RAPD marker to amplify a 436 bp fragment in the genomic DNA of female plants. This primer pair amplified the expected size of DNA fragment in female plants and four monoecious individuals collected from a natural population. The SCAR marker identified in this study can be used to distinguish female-flowering individuals at the seedling stage.

Bandsharing Values and Genetic Distances of Two Wild Shortnecked Clam, Ruditapes philippinarum Populations from the Yellow Sea Assessed by Random Amplified Polymorphic DNAs-Polymerase Chain Reaction

  • Yoon, Jong-Man;Kim, Yong-Ho
    • Journal of Aquaculture
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    • v.17 no.1
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    • pp.12-23
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    • 2004
  • Genomic DNAs were extracted from the muscle of twenty-two specimens of two shortnecked clam, Ruditapes phifippinarum populations collected in Anmyeondo and Seocheon. Genetic differences within and between populations were analysed by random amplified polymorphic DNAs-polymerase chain reaction (RAPD-PCR) using twenty arbitrary decamer primers. Out of 20 primers, 6 generated a total of 1,111 major and minor RAPD bands from individuals of two sites, producing approximately 4.2 average polymorphic bands per primer in individuals from Anmyeondo and ranging in size from less than 50 to larger than 1,500 base pairs (bp). The electrophoretic analysis of RAPD products amplified showed moderate levels of similarity among the different individuals in Seo-cheon population. The average bandsharing values (BS value) of the samples within population from Anmyeondo ranged from 0.155 to 0.684, whereas it was 0.143∼0.782 within population from Seocheon. The average BS value between individuals No. 13 and No. 14 from Seocheon was 0.782 which was higher than that of those from Anmyeondo. The single linkage dendrogram resulted from three primers (OPA-08, -09 and -20), indicating six genetic groupings composed of group 1 (No.4, 8 and 10), group 2 (No. 18), group 3 (No.2, 5 and 7), group 4 (No. 1, 3, 6, 9, 11, 12, 13, 14, 15 and 17), group 5 (16, 19 and 20) and group 6 (No. 21 and 22). In the Seocheon population, the individual No. 18 clustered distinctly from the others of this population. The observed genetic distance between the two populations from Anmyeondo and Seocheon was more than 0.209 (0.247 and 0.275). The shortest genetic distance (0.094) displaying significant molecular differences was between individuals No. 13 and No. 14. Especially, the genetic distance between individuals No. 22 and the remnants among individuals in two geographical populations was highest (0.275). This result illustrated that individual No.22 is distinct from other individuals within two shortnecked populations. The different geographical features of two sites may have caused the genetic diversity in two shortnecked clam populations.

Analysis of Genetic Relationship Among Native Pears Grown in Korea and Several Commercially Developed Cultivars from Two Pyrus Species Based on RAPD Analysis

  • Cho, Dong-Wook;Oh, Jin-Pyo;Chung, Kyu-Hwan
    • Korean Journal of Plant Resources
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    • v.20 no.6
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    • pp.563-569
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    • 2007
  • RAPD analysis showed that all the OTUs of 'Sandolbae' were the same species because amplified band patterns of all samples generated by each of 5 random primers were identical. Even though there were different environmental conditions, all the "Chuiangne" trees from three different places were the same species, and also all the "Cheongshilli" trees were the same species too. No genetic variations were detected between native Korean pears grown in the habitats and in the research field. Because 212 polymorphic bands were generated by 9 primers selected through primer screening, they were possible to analyze genetic relationship among naturally growing three native Korean pears and nine cultivars of Pyrus pyrifolia and P. communis. Based on the RAPD analysis, three main groups were formed. The first group represented the Six P. pyrifoia cultivars, the second group was the three native Korean pears, and the last group was the three P. communis cultivars. Genetic distance between 'Wonwhang' and 'Chojuro' was closer than other cultivars in group 1 since dissimilarity index value between these two cultivars was 50.82. However, genetic distance between 'Niitaka' and 'Chojuro' was the most distant compared to the others in group 1. In group 2, 'Sandlobae' was genetically closer to 'Chuiangne' than 'Cheongshilli' because dissimilarity index value between 'Sandlobae' and 'Chuiangne' was smaller, 50.82, than the value between 'Sandlobae' and 'Cheongshilli', 63.636. In group 3, 'Old Home' was genetically closer to 'Bartlett' than 'Kaiser Alexander(or Bosc)'. Group 3 composed of P. communis cultivars was genetically further than other two groups, P. pyrifolia cultivars and native Korean pears.

Distinction between Cold-sensitive and -tolerant Jute by DNA Polymorphisms

  • Hossain, Mohammad Belayat;Awal, Aleya;Rahman, Mohammad Aminur;Haque, Samiul;Khan, Haseena
    • BMB Reports
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    • v.36 no.5
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    • pp.427-432
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    • 2003
  • Jute is the principal coarse fiber for commercial production and use in Bangladesh. Therefore, the development of a high-yielding and environmental-stress tolerant jute variety would be beneficial for the agro economy of Bangladesh. Two molecular fingerprinting techniques, random-amplified polymorphic DNA (RAPD) and amplified-fragment length polymorphism (AFLP) were applied on six jute samples. Two of them were cold-sensitive varieties and the remaining four were cold-tolerant accessions. RAPD and AFLP fingerprints were employed to generate polymorphism between the cold-sensitive varieties and cold-tolerant accessions because of their simplicity, and also because there is no available sequence information on jute. RAPD data were obtained by using 30 arbitrary oligonucleotide primers. Five primers were found to give polymorphism between the varieties that were tested. AFLP fingerprints were generated using 25 combinations of selective-amplification primers. Eight primer combinations gave the best results with 93 polymorphic fragments, and they were able to discriminate the two cold-sensitive and four cold-tolerant jute populations. A cluster analysis, based on the RAPD and AFLP fingerprint data, showed the population-specific grouping of individuals. This information could be useful later in marker-aided selection between the cold-sensitive varieties and cold-tolerant jute accessions.

Induction of Valiant of Cyrtomium caryoptideum var. coreanum Nakai by Chemical Mutagenesis In vitro and RAPD Analysis (기내에서 화학돌연변이원 처리에 의한 참쇠고비의 변이주 유기 및 RAPD 분석)

  • Jeong Jin-A;Lee Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.19 no.2
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    • pp.374-380
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    • 2006
  • With the aim of inducing mutation in fern Cyrtomium caryoptideum var. coreanum, rhizome segments of In vitro-grown cultures were treated with chemical mutagens such as EMS, NMU and colchicine. Based on regeneration ratios, sensitivities for each treatments were assessed and also optimum treatment condition of each mutagens was explored. Optimum concentration for EMS treatment was considered to be 20 to 50mM and for NMU 5 to 10mM. NMU was found to be more effective in inducing chlorophyll and morphological variations than EMS. The RAPD were performed to check the genetic modification of phenotypical variants. As a result, polymorphic DNA band patterns between wild type and variants were observed by two 10-mer primers.

A RAPD-PCR Method for the Rapid Detection of Bacillus cereus

  • Lee, Ji-Yeon;Kwon, Gun-Hee;Park, Jae-Yong;Park, Cheon-Seok;Kwon, Dae-Young;Lim, Jin-Kyu;Kim, Jong-Sang;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.3
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    • pp.274-276
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    • 2011
  • Distinction of Bacillus cereus from other closely related bacilli is challenging and new efficient methods are continually demanded. From our previous work on RAPD profiles of bacilli, we found a possibility that B. cereus strains could be distinguished from other bacilli. In this work, RAPD-PCR profiles of B. cereus strains were obtained using a 10-mer (S30) as a primer, and a B. cereus specific 0.91-kb band was produced from all tested strains. The RAPD-PCR procedure also successfully detected B. cereus from spiked cheonggukjang when B. cereus cells were present at more than $10^2$/g sample.

Molecular Discrimination of Dinoflagellates Cochlodinium Polykrikoides Margalef, Gyrodinium Impudicum Fraga et Bravo and Gymnodinium Catenatum Graham using RAPD-PCR Method (RAPD-PCR 방법을 이용한 Cochlodinium polykrikoides Gyrodinium impudicum, Gymnodinium catenatum의 분자생물학적 진단)

  • Cho, Eun-Seob
    • Journal of Life Science
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    • v.13 no.5
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    • pp.651-657
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    • 2003
  • Randomly amplified polymorphic DNA (RAPD) analysis was used to study genetic relationships among C. polykrikoides, G. impudicum and G. catenatum, which possess similar morphological features. Four of 12 primers were selected and 59 amplification products ranged from 0.2 kb to 3.0 kb. The number of polymorphic products in C. polykrikoides, G. impudicum and G. catenatum was 16 (27.1%), 8 (13.5%), and 16 (27.1%), respectively, while 17 were monomorphic. Number of species-specific bounds was 26 (44.0%), 34 (57.6%), 26 (44.0%) in C. polykrikoides, G. impudicum and G. catenatum, respectively. The genetic similarity between C. polykrikoides and G. impudicum/G. catenatum was 0.83, whereas G. impudicum and G. catenatum was 0.78. Our results suggest that C. polykrikoides, G. impudicum and G. catenatum are extremely different on the basis of RAPD analysis, despite similarity based on their morphology. The RAPD technique appears to be efficient in detecting genetic variation in these dinoflagellates.

Isolation and Identifieation of Entomopathogenic Nematodes from Soil and Insect (토양과 곤충 사체로부터 곤충병원성 선충의 분리 및 동정)

  • 한상미;한명세
    • Korean Journal of Environmental Biology
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    • v.17 no.3
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    • pp.321-330
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    • 1999
  • Nematodes were isolated using silkwom trap through the investigation of 100 soil samples from various biotopes in Korea. The 30 nematode strains from soil and dead insects by the pathogenicity aganinst silkworms (Bombyx mori mori) and insect pests of Calliphora vomitoria, Pseufazetia separata, Palomena angulosa, and Melolontha incana. Mortailty of the silkworm larvae and pupae were as high as 100% by nematode infection, those of insect of pests were varied from 20 to 100%. The 30 strains of entemopathogenic nematodes were classified into five groups of Rhabditidae, Diplogatroidae, Heterorhabitidae, Steinernematidae, and Tylenchida by morphological criteria. The genetic relationships among the 30 nematode strains were analyzed by various RAPD bands with twenty primers. The 30 nematode strains were classified into six major subgroups on the basis of the genetic similarity coefficient of 0.853. The grouping by RAPD was agree with those of morphological taxa in discrimination of the higher group, however, was not completely agree in the subgroup. The family Steinernematidae belong to Rhabditida was clarified as closer to the Tylenchida, rather than the other Rhabditida of Heterorhabitidae, Rhabditidae, and Diplogatroidae in genetic distance valule. From the result of the morphological classification and RAPD of the genomic DNA showed that genetic relationship analysis furnish infurmation on phylogenetic classification and relationships of entomopathogenic nematodes. The application of genetic similarity will overcome the limitation of taxonomy and classification of morphologically simple nematode. Several primers were confirmed those utility of identification for individual nematode strains, the methods of molecular genetics secured the simplicity, rapidity and accuracy on the selection of entomopathogenic nematodes.

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