• Title/Summary/Keyword: RAPD markers

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Relationship of Lycoris (Amaryllidaceae) Based on RAPD Markers (RAPD markers에 의한 상사화속 식물의 유연관계)

  • Tae, kyoung-Hwan;Kim, Yong Hyun;Shin, Young-Hwa;Kang, Shin-Ho;Kim, Joo-Hwan;Ko, Sung-Chul
    • Korean Journal of Plant Taxonomy
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    • v.38 no.1
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    • pp.17-29
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    • 2008
  • Phylogenetic relationships were examined for 17 taxa of Lycoris by RAPD analysis. The length of the amplified DNA fragments ranged from 300 bp to 1,700 bp. 57 scorable RAPD markers were observed from PCR reactions with five random oligoprimers. The analysis by UPGMA sepatated the examined taxa of Lycoris into were clusters. First group was comprised of ten taxa of L. chinensis var. sinuolata, L. sanguinea var. koreana, L. uydoensis, L. flavescens, L. radiata var. pumila, L. radiata, L. squamigera, L. chejuensis, L. aurea and L. guangxiensis, second group of L. haywardii, L. sprengeri, L. rosea, L. straminea and L. houdyshii, third group of outgroup of Narcissus tazetta var. chinensis and Crinum asiaticum var. japonicum. From the viewpoint of cytological characters such as polyploidy and karyotype, the RAPD analysis was very useful to show the relationship among the intraspecific taxa of Lycoris.

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.

Confirmation of $F_1$ Hybridity Using RAPD Markers in Soybean

  • Chung, Jong-Il;Ko, Mi-Suk;Shim, Jung-Hyun;Kim, Seok-Hyeon;Kang, Jin-Ho
    • Plant Resources
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    • v.2 no.1
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    • pp.22-25
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    • 1999
  • Molecular markers are useful to confirm the hybridity of F1 plant derived from cross of two homozygous parents with similar morphological traits. RAPD markers were used to test F1 hybrid plant obtained from cross of two homozygous soybean (Glycine max) parents. Fl plant for cross I was made from the mating of Hobbit87 (female) and L63-1889 (male) and Fl plant for cross II was obtained from the mating of H1053 (female) and L63-1889 (male). Selfing plant per each cross was also obtained. Among 20 Operon primers used, OPA04 and OPA09 show polymorphism between cross I and II parent. Band in size 1Kb of OPA04 and 2.1Kb of OPA09 primer was polymorphic band. This fragment identified Fl hybrid plant and selfing plant in cross I and II. Female parent Hobbit87 in cross I and H1053 in cross II has no this fragment (recessive allele). However, male parent L63-1889 and Fl hybrid plant in cross I and II has this size of polymorphic band (dominant allele). This indicated that Fl hybrid and selfing plants were detected by RAPD marker before phenotypic marker would be used to identify Fl hybridity. Amplification products of selfing plant for cross I and II were completely same to the those of female parent. When mature, flower color of Fl hybrid plant in cross I and II was purple and flower color of selfing plant in cross I and II was white. Purple flower is dominant trait. Fl hybridity was successfully detected at very early growth stage using RAPD marker. Therefore, RAPD marker can be used broadly to confirm Fl hybridity in many crops.

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RAPD Analysis for Genetic Diversity of Melon Species (참외와 멜론의 유전적 다양성에 대한 RAPD 분석)

  • Mo, Suk-Youn;Im, Sung-Hee;Go, Gwan-DaI;Ann, Chong-Mun;Kim, Doo Hwan
    • Horticultural Science & Technology
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    • v.16 no.1
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    • pp.21-24
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    • 1998
  • RAPD markers were analyzed in order to detect the genetic variation and diversity of the fifty-two melon lines. SDS extraction method produced more and purer DNA than CTAB method. RAPD reaction conditions were optimized as follows ; 10ng template DNA, 270nM primer, $200{\mu}M$ each of dATP, dCTP, dGTP and dTTP, $0.3{\mu}unit$ dynazyme and 10x buffer brought to $15{\mu}l$ final volume with distilled water. The adequate annealing temperature was $39^{\circ}C$ and forty cycles of amplification produced the best RAPD band patterns. Among a total of 123 bands from 12 random primers, 25 polymorphic bands(20%) were selected as reliable markers. The average number of polymorphic bands per primer was 2.1 among the 52 lines. Intragroup genetic relationship based on the marker difference was closer than intergroup genetic relationship. The 52 lines could be grouped into two major group (Korean landraces and melon lines) and then melon group subdivided into two subgroups (net melon lines and no-net melon). This result corresponded to morphological grouping. Eight RAPD markers separated the Korean landraces and melon groups and four RAPD markers separated net melon and no-net melon groups.

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RAPD-SCAR Markers Linked to Medium-Leaf Zoysiagrass Ecotypes (한국잔디 중지 변이개체와 연관된 RAPD-SCAR 마커)

  • Chung, Sung Jin;Park, Su Jeong;Kim, Hun Joong;Yang, Geun-Mo;Choi, Joon-Soo;Oh, Chan-Jin;Jang, Deok-Hwan;Song, In-Ja;Lee, Geung-Joo
    • Weed & Turfgrass Science
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    • v.2 no.2
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    • pp.191-197
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    • 2013
  • Two medium-leaf ecotypes (CY6069, CY6097) belonging to one species (Zoysia japonica) of Korean lawngrasses were selected in sod production fields in Jang Seong, Korea. They were reported to have distinct morphological and growth rate characteristics different from the preferred medium-leaf type zoysiagrass in Korea. This study was conducted to define further the genotypic difference at the molecular level and to develop DNA marker based on the specific DNA fragment. Polymorphic DNA fragments were first explored by using randomly amplified polymorphic DNA (RAPD) primers, which were then converted into PCR-based sequence characterized amplified region (SCAR) markers. The CY6069-specific primer set amplified about 550 bp successfully, while the CY6097 marker produced the expected 690 bp band, by which those markers were nominated by CY6069_550 and CY6069_690 SCARs, respectively. Together with the reported morphological and other phenotypic features, the SCAR markers confirmed in this study will be useful to identify those medium-leaf zoysiagrass genotypes when they are cultivated with other vegetatively propagated warm-season turfgrasses in sod farms.

Discrimination of Korean Apple Cultivars Using Combination of RAPD-SCAR Markers (RAPD-SCAR 마커 조합을 이용한 국내 육성 사과 품종 판별)

  • Cho, Kang-Hee;Heo, Seong;Kim, Hyun-Ran;Kim, Jeong-Hee;Shin, Il-Sheob;Han, Sang-Eun;Kim, Se-Hee;Kim, Dae-Hyun
    • Horticultural Science & Technology
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    • v.28 no.5
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    • pp.828-835
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    • 2010
  • Conventional methods for identification of apple cultivars are based on the evaluation of sets of morphological characteristics, however, closely related cultivars often cannot be distinguished by morphological traits. This study was conducted to develop DNA markers for discrimination of the apple cultivars bred in Korea. Thirty random primers generated eighty-three random amplified polymorphic DNA (RAPD) markers from thirty-one Korean bred and introduced apple cultivars. Fifty-two RAPD fragments were cloned and sequenced for conversion into sequence characterized amplified region (SCAR) markers. Among them only seventeen SCAR markers resulted in the amplification of single major bands the same size as the RAPD fragment cloned. Several combinations of six (AN11_433, AN08_566, A408_592, AK17_653, AO04_711, AO04_779 or AW15_368, AN11_433, A408_592, AK17_653, AO04_711, AO04_779, or AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AO04_779) to seven (AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AM16_708, AO04_779 or A330_424, AN11_433, AG14_502, AN08_566, A408_592, AK17_653, AO04_779 or A330_424, AN11_433, AK14_564, A408_592, AK17_653, AM16_708, AT14_789) SCAR markers provided enough polymorphism to identify sixteen Korean apple cultivars among thirty-one tested cultivars. Therefore, application of the seventeen SCAR markers was sufficient to identify the thirty-one tested apple cultivars. These markers could be utilized as a reliable tool for cultivar discrimination of Korean apples.

Analysis of Genetic Variation in Botrytis cinerea Isolates Using Random Amplified Polymorphic DNA Markers

  • Choi, In-Sil;Kim, Dae-Hyuk;Lee, Chang-Won;Kim, Jae-Won;Chung, Young-Ryun
    • Journal of Microbiology and Biotechnology
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    • v.8 no.5
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    • pp.490-496
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    • 1998
  • Random amplified polymorphic DNA (RAPD) markers were used to survey genetic variability among 34 Botrytis cinerea isolates from nine different host plants in Korea. For RAPD analysis, 115 arbitrary decamer primers were initially screened for polymorphic major DNA bands with 11 representative B. cinerea isolates. Eleven primers that initially detected polymorphisms were tested a second time with additional 23 isolates of B. cinerea as well as one isolate of Botrytis squamosa as an outgroup. The RAPD analyses revealed that all isolates except one showed different molecular phenotypes. Dendrograms obtained from dissimilarity matrices using the unweighted paired group method of arithmetic means (UPGMA) showed the 36.4% to 90.0% similarity among all B. cinerea isolates. The B. squamosa isolate showed the least similarity to all B. cinerea isolates. The cluster analyses indicated no correlation among all the characteristics examined including molecular phenotypes, host and geographic origins, year of isolation, or pathogenicity. The RAPD data suggest that a high level of genetic variation exists among Korean populations of B. cinerea and it seems to be caused by heterokaryosis among preexisting molecular phenotypes.

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Genetic Diversity and Differentiation of Colletotrichum spp. Isolates Associated with Leguminosae Using Multigene Loci, RAPD and ISSR

  • Mahmodi, Farshid;Kadir, J.B.;Puteh, A.;Pourdad, S.S.;Nasehi, A.;Soleimani, N.
    • The Plant Pathology Journal
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    • v.30 no.1
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    • pp.10-24
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    • 2014
  • Genetic diversity and differentiation of 50 Colletotrichum spp. isolates from legume crops studied through multigene loci, RAPD and ISSR analysis. DNA sequence comparisons by six genes (ITS, ACT, Tub2, CHS-1, GAPDH, and HIS3) verified species identity of C. truncatum, C. dematium and C. gloeosporiodes and identity C. capsici as a synonym of C. truncatum. Based on the matrix distance analysis of multigene sequences, the Colletotrichum species showed diverse degrees of intera and interspecific divergence (0.0 to 1.4%) and (15.5-19.9), respectively. A multilocus molecular phylogenetic analysis clustered Colletotrichum spp. isolates into 3 well-defined clades, representing three distinct species; C. truncatum, C. dematium and C. gloeosporioides. The ISSR and RAPD and cluster analysis exhibited a high degree of variability among different isolates and permitted the grouping of isolates of Colletotrichum spp. into three distinct clusters. Distinct populations of Colletotrichum spp. isolates were genetically in accordance with host specificity and inconsistent with geographical origins. The large population of C. truncatum showed greater amounts of genetic diversity than smaller populations of C. dematium and C. gloeosporioides species. Results of ISSR and RAPD markers were congruent, but the effective maker ratio and the number of private alleles were greater in ISSR markers.

The Genetic Relationship of Zelkova Serrata Registered s the Natural Monument Using RAPD Markers (RAPD marker로 추적한 천연기념물로 지정된 느티나무의 유연관계)

  • 강경홍;정영재;김홍남
    • Korean Journal of Environmental Biology
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    • v.17 no.1
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    • pp.89-94
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    • 1999
  • The genetic polymorphism and relationship among 14 Zelkova serrata registered as the natural monument in Korea were investigated using RAPD markers. N-J tree indicated that individuals in Kangwon-do were clustered and those in Yangam-gun and Damyang-gun of Chollanam-do were done closely. However, all the others were not agreed with the geographical distribution. Maybe this discordance was resulted from their movement by human being than any biological factors. The polymorphic percentages among individuals were from 77.8 to 100. From the high polymorphism, it was supposed that existing natural monuments were independently originated in the ancestors of long differentiated populations.

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RAPD marker variations between and within the species of Korean Suaeda (한국산 나문재속의 종내·종간 RAPD marker 변이)

  • Shim, Hyun-Bo;Choi, Byoung-Hee
    • Korean Journal of Plant Taxonomy
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    • v.34 no.1
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    • pp.63-74
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    • 2004
  • The genus Suaeda is one of the most popular plants on salt marsh areas in Korean Peninsular. The entities that comprise taxa in Korea exhibit widely overlapping ranges in all morphological attributes. Ramdom amplified polimorphic DNA (RAPD) markers were used to clarify taxonomic delimitation among the Korean taxa of Suaeda and to analyse genetic variations among the populations of S. japoniro in western and southern coastal regions. Six decamer primers amplified a total of 65 scorable bands, of which 61 were polymorphic. In all primers investigated, S. glauro of sect. Schanginia is the most distinctive species, compared with others of sect. Heterosperma. S. japonica, S. maritima, and S. malacosperma, which have been hardly distinguished each other by external morphology, were readily recognized by its specific DNA bands. The characteristic RAPD markers were identified local populations of S. japonica, but this feature was not revealed within population.