• Title/Summary/Keyword: RAPD PCR

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분자생물학적 기법을 사용한 질산화유도 반응기내 군집동태변화

  • Jo, Sun-Ja;Jeong, Yong-Ju;Kim, Jeong-Cheol;Lee, Sang-Jun
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2005.05a
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    • pp.247-248
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    • 2005
  • 본 연구는 서로 다른 조건에서 질산화를 유도한 반응기의 군집동태를 살피기 위해 RAPD, ARDRA, DGGE와 같은 기법들을 적용시켜 반응기 초기의 슬러지 구성 군집의 상태 와 질산화 유도후의 군집 변화를 살펴보고자 하였다. 결과적으로 1.5 kb정도의 165 rDNA를 이용한 RAPD와 ARDRA에서는 RAPD에 의한 군집 Patterns변화가 훨씬 다양했으며, 250 bP정도의 PCR산물로 분리를 시도한 DGGE에서도 비교적 예상했던 바와 같이 군집이 단순화되는 양상을 볼 수 있었다.

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Comparison of Pepper Anthracnose Pathogens from Korea and China

  • Shin, Hyun-Joo;Chen, Zhu-Jun;Hwang, Jae-Moon;Lee, Soon-Gu
    • The Plant Pathology Journal
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    • v.15 no.6
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    • pp.323-329
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    • 1999
  • Anthracnose of pepper casued by Colletotrichum spphas been a great problems for pepper production in Korea and China. Especially Colletotrichum gloeosporioides was found predominantly over cultivation areas during infection periods and caused severe rots on bath unripe and ripe fruits that resulted in major yield losses. In this study, comparison of Colletrichum spp.isolated from Korea and China in morphology and pathgenicity, and RAPD-PCR analysis were conducted. Based on morphological characteristics, the pathogen isolates, K1 and C1, K2 and C2, and K3 and C3 were identified as Colletotrichum gloeosporioides (G) type, C. gloeosporioides (R) type and C. coccodes, respectively. in pathogenicity test, K1 and C1, and K2 and C2 were found to attack mainly fruits and to be the most virulent among isolates. K3 and C3 were strongly virulent to leaves and seedling. Pathogenicity between Korean and Chinese isolates. K3 and C3 were strongly virulent to leaves and seedling. Pathogenicity between Korean and Chinese isolates did not show any difference. Results of the RAPD-PCR analyses indicate the varying levels of molecular diversity within and between Colletotrichum spp.of Korea and China. The similarities between K1 and C1, K2 and C2, and K3 and C3 were 85.71%, 71.43% and 50.0% respectively.

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Analysis of Genetic Relatedness by Random Amplified Polymorphic DNA (RAPD) in Pecan Taxa (RAPD를 이용한 Pecan 품종의 유전적 관계 분석)

  • 신동영;김회택;박종인;노일섭
    • Korean Journal of Plant Resources
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    • v.13 no.1
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    • pp.1-10
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    • 2000
  • Pecan is deciduous tree and belongs to the Julandaceae family. Pecan is an economically important as a nut and timber crop. Heterozygosity is expected to be high for typically cross-pollinated. Yet little is known about the nature of genetic variation within this species. In addition, the pedigree of many pecan cultivars remains unknown or is questionable. In this study, the phylogenetic relationships between 22 pecan cultivars and its analyzed by RAPD (randomly amplified polymorphic DNA). PCR Amplification used 40 randomly selected oligoes as primers. Based on their genetic similarities derived from the RAPD data, the 22 pecan cultivars were classified into different five groups in agarose gel. The 22 pecan cultivars were classified into five sectional groups by UPGMA clustering analysis, too. C. flacra and Black walnut showed the 0.9 of similarity index and Farley, Pawnee showed the 0.85 of similarity index. The 22 pecan cultivars were classified into different five groups by analysis of the 4% polyacrylamide gel fraction. (Group I : 1, 2, 3, 4, 13, 16, 17, 20, 21 Group II : 14,18 GroupIII : 6,12 GroupIV : 5, 11, 15, 19, 22 CroupV : 7, 8, 9, 10) Group V show the 1.0 of similarity index and Farley, Sturya, Clarke, Pawnee show the 0.98 of similarity index and Kiowa, Schley show the 0.92 of similarity index. Results from this study indicated that RAPD can be used to establish the genetic relationships among the 22 pecan cultivars. Similarity coefficients generally agreed with what would be predicted in cultivars with known pedigrees, and we could accurately construct relationships among cultivars. In addition, we have shown that RAPD provides useful information on the origin of unknown cultivars.

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Development of SCAR Markers for the Discrimination of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma based on the RAPD (RAPD 분석을 통한 대황(大黃)과 종대황(種大黃) 감별용 SCAR 유전자 마커 개발)

  • Moon, Byeong-Cheol;Lee, Young-Mi;Chun, Jin-Mi;Lee, A-Young;Yoon, Tae-Sook;Cheon, Myeong-Sook;Choo, Byung-Kil;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.115-120
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    • 2009
  • Objectives : Due to the morphological similarity and frequent occurrence of intermediate forms as well as morphological variations of aerial part, the correct identification between Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma is very difficult. To develop a reliable method for correct identification and improving the quality standards of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma, we analyzed RAPD and developed SCAR marker. Methods : To amplify target DNA at the genomic level, 32 Operon 10-mer random primers were applied with four Rheum species, R. officinale, R. palmatum, R. tanguticum and R. undulatum. The nucleotide sequences were determined and species-specific primers were prepared depending on the species-specific RAPD amplicons after subcloned into the pGEM-Teasy vector. To develop the SCAR markers, species-specific PCR amplification and multiplex-PCR were carried out using the single species-specific primer pairs and combinations of them, respectively. Results : We used RAPD analysis of four Rheum plant species to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed two SCAR markers that amplified 314 bp and 390 bp DNA fragments in only R. undulatum but not in R. officinale, R. palmatum, R. tanguticum and R. undulatum, for distinguishing Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. Conclusions : These genetic markers can be used for the efficient discrimination of plants species and commercial herbal medicines between Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma, to ultimately prevent indiscriminate distribution and prescription of these herbal medicines.

Genetic Diversity of Orobanche cumana Populations in Serbia

  • Ivanovic, Zarko;Marisavljevic, Dragana;Marinkovic, Radovan;Mitrovic, Petar;Blagojevic, Jovana;Nikolic, Ivan;Pavlovic, Danijela
    • The Plant Pathology Journal
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    • v.37 no.6
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    • pp.512-520
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    • 2021
  • In this study, we report genetic characterization of Orobanche cumana, the causal agent of sunflower wilting in Serbia. The genetic diversity of this parasitic plant in Serbia was not studied before. Random amplified polymorphic DNA (RAPD) markers and partial rbcL gene sequences analysis were used to characterize the O. cumana populations at the molecular level. While phylogenetic analyses of RAPD-PCR amplicons were performed using unweighted pair-group Method analyses, rbcL gene sequences were analyzed using neigbor joining method and minimum spanning tree. Molecular analyses of RAPD-PCR analysis revealed high genetic diversity of O. cumana populations which indicated high adaptive potential of this parasitic weed in Serbia. Further analyses of rbcL gene using minimum spanning tree revealed clear differences among diverse sections of Orobanche genus. Although this molecular marker lacked the resolution to display intrapopulation diversity it could be a useful tool for understanding the evolution of this parasitic plant. Our results suggested that O. cumana has great genetic potential which can lead to differentiation of more virulent races which is important for determining crop breeding strategies for their control.

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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Diversity of I-SSR Variants in Gingko biloba L. Planted in 6 Regions of Korea (국내(國內) 6개(個) 은행(銀杏)나무 식재지(植栽地)에 있어서 I-SSR 변이체(變異體)의 다양성(多樣性))

  • Hong, Yong-Pyo;Cho, Kyung-Jin;Hong, Kyung-Nak;Shin, Eun-Myeong
    • Journal of Korean Society of Forest Science
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    • v.90 no.2
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    • pp.169-175
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    • 2001
  • Genomic DNAs were extracted from the leaves of 182 ginkgo trees (Ginkgo biloba L.) planted in 6 regions and subjected to the analysis of both I-SSR and RAPD markers. A total of 227 amplicon variants were generated by PCR using 15 I-SSR primers and 67 amplicons by PCR with 5 RAPD primers. Levels of genetic diversity within 6 populations were turned out to be similar (Shannon's Index, I-SSR : 0.35~0.40; mean of 0.38, RAPD : 0.31~0.38; mean of 0.35, combined : 0.35~0.40; mean of 0.37). Ranks of the level of genetic diversity estimated from I-SSR, RAPD, and combined data were not coincided each other. Majority of genetic diversity was allocated among individuals within populations (I-SSR : 94.31%, RAPD : 93.62%, combined : 93.57%), which resulted in pretty low level of population differentiation. Genetic differentiation between male and female groups was turned out to be quite low (I-SSR : 0.03, RAPD : 0.091, combined : 0.043), which slightly fluctuated when analysis was restricted to the data obtained from 3 regions where both male and female trees were sampled (I-SSR : 0.038, RAPD : 0.084, combined : 0.047). Genetic relationships among the populations, reconstructed by UPGMA, were not coincided with geographic affinity, which might be resulted from sharing of seed sources in some regions. Whereas independent cluster analyses with I-SSR data and RAPD data, respectively, reclassified by sexes revealed two sexual groups in which all the male and the female populations were clustered together, cluster analysis with combined data did not show clear sexual grouping.

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Construction of a Genetic Linkage Map of Shiitake Mushroom Lentinula Edodes Strain L-54

  • Hoi-Shan, Kwan;Hai-Lou, Xu
    • BMB Reports
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    • v.35 no.5
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    • pp.465-471
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    • 2002
  • From fruiting bodies of L. edodes strain L-54, single-spore isolates (SSIs) were collected. Two parental types of L-54 were regenerated via monokaryotization. By means of random-amplified polymorphic DNA (RAPD), DNA samples from L-54, its two parental types, and 32 SSIs were amplified with arbitrary primers. Dedikaryotization was demonstrated, and 91 RAPD-based molecular markers were generated. RAPD markers that were segregated at a 1:1 ratio were used to construct a linkage map of L. edodes. This RAPD-linkage map greatly enhanced the mapping of other inheritable and stable markers [such as those that are linked to a phenotype (the mating type), a known gene (priA) and a sequenced DNA fragment (MAT)] with the aid of mating tests, bulked-segregant analysis, and PCR-single-strand conformational polymorphism. These markers comprised a genetic map of L. edodes with 14 linkage groups and a total length of 622.4 cM.

Mendelian Inheritance of Inter-Simple Sequence Repeats Markers in Abies Koreans Wilson (구상나무에 있어서 Inter-Simple Sequence Repeats Marker의 유전양식(遺傳樣式))

  • Hong, Yong-Pyo;Cho, Kyung-Jin;Kim, Yong-Yul;Shin, Eun-Kyeong
    • Journal of Korean Society of Forest Science
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    • v.87 no.3
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    • pp.422-428
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    • 1998
  • Polymerase chain reaction(PCR)-based inter-simple sequence repeats(I-SSR) markers were analyzed in 48 megagametophytes of a single tree of Abies koreana $W_{ILS}$. Nineteen of the 35 primers, screened with 6 megagametophyte DNA and produced the clearest amplification products in the preliminary experiment, were used for PCR with 48 megagametophyte DNAs sampled from a single tree. On the basis of the chi-square test, a total of 51 amplicons, amplified by the 19 primers, were revealed to be segregated according to the Mendelian ratio(i.e., 1 : 1 segregation ratio) in the 48 megagametophytes at 5% significance level. Based on the linkage analysis, the observed 51 Mendelian loci turned out to be unlinked each other, which suggested that they are evenly distributed in the genome. However, majority of RAPD markers are known to belong to the independent linkage blocks, which frequently results in the amplification of RAPD markers from the restricted regions of the genome. Owing to the nature of even distribution of the 51 loci observed in this study, the I-SSR markers could give better resolution of estimating genetic diversity from the whole genome than RAPD markers. And I-SSR markers are also more suitable than RAPD markers for reconstructing phylogenetic relationship by a cladistic method which requires to fulfil the assumption of independent evolution of the different characters.

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Genetic Relationship among Ostericum koreanum Kitakawa Collections by RAPD Analysis (RAPD에 의한 강활 수집종간의 유연관계 분석)

  • Kim, Soo-Yong;Sim, Yong-Gu;Kwon, Soon-Tae;Oh, Sei-Myung
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.3
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    • pp.109-113
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    • 2005
  • To analyze the genetic relationship 8 accessions of Ostericum koreanum Kitakawa, random amplified polymorphic DNA(RAPD) analysis was performed using 60 Operon primers. The 25 primers out of 60 random primers were amplified DNA by PCR using genomic DNA of O. koreanum. Eighty-five (49.1%) among 173 bands derived from 25 primers showed poly morphism, On the basis of similarity coefficient analysis by UPGMA, 8 accessions of O. koreanum Kitakawa could be classified into three groups at the similarity coefficient value of 0.71. Group I contained three accessions (Nam Gangwhal), Group II contained one accession (Nam Gangwhal) and Group III contained four accessions (Buk Gangwhal), The range of total genetic similarity coefficient value of 8 accessions of O. koreanum Kitakawa was $0.63{\sim}0.96$. Buk Gangwhal was flowered 18 to 26 days earlier than Nam Gangwhal, and Nam Gangwhal leaf stalk was thin and long as bolting rate high compared with Buk Gangwhal.