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

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Comparison of Cellular Fatty Acid Composition and Genotypic Analysis of Bifidobacterium longum MK-G7 with Commercial Bifidobacteria Strains

  • Jung, Hoo-Kil;Kim, Eung-Ryool;Ji, Geun-Eog;Park, Jong-Hyun;Cha, Seong-Kwan;Juhn, Suk-Lak
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.143-146
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    • 2000
  • This study was conducted to compare the cellular fatty acid composition and genotypic analysis of Bifidobacterium longum MK-G7 originated from Koreans with other commercial type strains of bifidobacteria. The cellular fatty acid of Bif. longum MK-G7 was shown to be composed of $C_{160FAME},C_{181\;c18DMA},C_{18.1\;CIS9\; FAME},C_{14.0FAME},C_{19\;0cye9,10 DMA},Feature7(C_{17.2 FAME), and Feature 10(C_{181\; Cll/t9/t6 FAME}$. Bif. longum MK-G7 showed 99.9% homology and the highest relatedness with Bif. longum ATCC 15707 type strain. Both Bif. longum MK-G7 and Bif. longum ATCC 15707 showed 153 bp products on RAPD (randomly amplified polymorphic DNA) analysis, however, they showed quite different band patterns on PFGE (pulsed-field gel electrophoresis) analysis. Consequently, our present study showed that Bif. longum MK-G7 was different from any commercial type strains of Bif. longum tested.

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Microscopical observation and randomly amplified polymorphic DNA (RAPD) analysis of artificially cultivated Ganoderma applanatum

  • Woo-Sik Jo;Young-Hyun Rew;Seung-Chun Park
    • Journal of Mushroom
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    • v.22 no.2
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    • pp.48-52
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    • 2024
  • This study investigated the microscopic characteristics and genetic relationships of Ganoderma applanatum fruiting bodies. Basidiospores were brown, ellipsoid, and had one or two large vacuoles and a double wall. The surface of basidiospores was smooth or wrinkled and most had numerous small and shallow holes. The length and width of basidiospores of Ganoderma applanatum isolates GBGA-01, GBGA-02, ASI 50167, ASI 52821, ASI 52822, ASI 52823, and ASI 53399 were on average 7.6×4.8 ㎛, 7.9×4.6 ㎛, 7.7×4.9 ㎛, 8.2×5.3 ㎛, 7.7×5.0 ㎛, 8.0×4.9 ㎛, and 7.9×4.9 mm, respectively. In contrast, the basidiospores of Ganoderma lucidum isolate ASI 7125 were 7.7×5.2 ㎛. Using the universal ITS1/ITS4 primer set, the ITS region of the isolates were amplified and sequenced. The ITS sequences were very closely related to G. applantum isolate GBGA-01, GBGA-02, ASI 50167, ASI 52821, ASI 52822, ASI 52823 and ASI 53399, but were not the same species. Whereas, G. lucidum isolate ASI 7125 belongs to different group.

Isozyme and Randomly Amplified Polymorphic DNA (RAPD) Analysis for Genetic Relationship among Calanthe discolor, C. sieboldii, and C. bicolor Native to Cheju Island (동위효소와 RAPD 법을 이용한 제주 자생 새우란, 금새우란, 왕새우란의 근연관계 분석)

  • Hyun, Myung-Ryuk;Choi, Ji-Yong;Suh, Jung-Nam;So, In-Sup;Lee, Jong-Suk
    • Horticultural Science & Technology
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    • v.17 no.2
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    • pp.141-143
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    • 1999
  • Genetic relationship of Calanthe discolor, C. sieboldii, and C. bicolor for elucidating the classification was investigated. Electrophoretic zymograms for either peroxidase or esterase isozymes indicated that bands of C. bicolor appear in the zone where those of C. discolor and C. sieboldii are located. Genetic relationship among the three Calanthe species using RAPD analysis showed that C. discolor and C. sieboldii are more distant each other than C. bicolor, demonstrating the genetic position of C. bicolor between the other two. It was assumed that C. bicolor is a natural hybrid between C. discolor and C. sieboldii.

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Phylogenetic Relationship of Ligularia Species Based on RAPD and ITS Sequences Analyses (RAPD 및 ITS 염기서열 분석을 이용한 곰취 속(Ligularia) 식물의 유연관계 분석)

  • Ahn, Soon-Young;Cho, Kwang-Soo;Yoo, Ki-Oug;Suh, Jong-Taek
    • Horticultural Science & Technology
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    • v.28 no.4
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    • pp.638-647
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    • 2010
  • The genetic relationships in 5 species of $Ligularia$ were investigated using RAPD (Randomly Amplified Polymorphic DNA) and ITS (Internal Transcribed Spacer) sequences analyses. In RAPD analysis, sixty three of 196 arbitrary primers showed polymorphism. The amplified fragments ranged from 0.2 to 1.6 kb in size. The dendrogram was constructed by the UPGMA clustering algorithm based on genetic similarity of RAPD markers. A total of 16 accessions were classified into 5 major groups corresponding each species at the similarity coefficient value of 0.77. In the ITS sequence analysis, the size of ITS 1 was varied from 248 to 256 bp, while ITS 2 was varied from 220 to 222 bp. The 5.8S coding region was 164 bp in lengths. Forty nine sites (10.2%) of the 478 nucleotides were variable, and the G+C content of ITS region ranged from 49.4 to 53.5%. In the ITS tree, five species of $Ligularia$ were monophyletic, and $L.$ $taquetii$ was the first branching within the clade. $Ligularia$ $intermedia$ formed a clade with $L.$ $fischeri$ var. $spiciformis$ (BS=79), and $L.$ $stenocephala$ and $L.$ $fischeri$ were also claded. Two data sets were congruent, except of the position of $L.$ $fischeri$ var. $spiciformis$.

Randomly Amplified Polymorphic DNA (RAPD) Analysis on Compatibility of Korean Native Cymbidium goeringii with Other Cymbidium Species (한국춘란(韓國春蘭)과 다른 Cymbidium간의 교배친화성(交配親和性)에 대한 RAPD 분석)

  • Choi, Ji-Yong;So, In-Sup;Pak, Chun-Ho;Kwack, Beyoung-Hwa
    • Horticultural Science & Technology
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    • v.16 no.3
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    • pp.361-363
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    • 1998
  • Cymbidium goeringii native to Korea and other orchid plants were pot-grown from spring to autumn under the greenhouse conditions, and were subjected to artificial pollination to elucidate the compatibility by revealing viable seed formation. A notable compatibility was found when Cym. goeringii was selfed and was crossed with either Cym. ensifolium, Cym. kanran, Cym. sinense, Cym. sinense for. albo-jucundissimum, Cym. 'Crystal Cherry Angel', or Cym. 'Anmitsu Hime'. Cym. goeringii, however, did not show such compatibility when crossed with either Cym. faberi, Cym. aloifolium, Dendrobium chrysotoxum, or Phalaenopsis spp. RAPD analysis indicated that taxa relationship between Cym. goeringii and either Cym. faberi or Cym. aloifolium (respective chromosome number, 2n=40) was distant, showing no compatibility, and even more distant in the case of cross-pollination between Cym. goeringii and either Dendrobium chrysotoxum or Phalaenopsis spp. having different chromosome number from all Cymbidium species.

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Population analysis of eelgrass, Zostera marina L. in Geojedo, Gaedo, and Jedo on the southern coastal water of Korea using RAPD-PCR (RAPD 방법을 이용한 거제도, 개도, 제도해역에서 채집한 말잘피 개체분석)

  • Cho, Eun-Seob;Lee, Sang-Yong;Kim, Jeong-Bae
    • Journal of Life Science
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    • v.17 no.4 s.84
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    • pp.455-461
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    • 2007
  • Assessments of population genetic structure and diversity can be of value in formulating management plans for threatened eelgrass(Zostera maim). Using randomly amplified polymorphic DNA markers, we found evidence of significant genetic structure among the populations of eelgrass sampled at three areas(Geojedo, Gaedo, and Jedo). A highly isolated(>100 km apart) population from the Geojedo had a long genetic distance(0.16), whereas the populations from the Gaedo and Jedo(<10 km apart) exhibited far less distance(0.08). The analysis of similarity within population showed that Geojedo was over 70%, which was of lower value than of Gaedo and Jedo. Based on these results, we realized that heterogeneous population was in accordance with geographic separation. This is caused by limited seed dispersal and interrupted gene flow, although the sample size is small.

An analysis of the genetic diversity of a riparian marginal species, Aristolochia contorta (수변 경계종인 쥐방울덩굴의 유전적 다양성 분석)

  • Nam, Bo Eun;Park, Hyun Jun;Son, Ga Yeon;Kim, Jae Geun
    • Journal of Wetlands Research
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    • v.22 no.2
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    • pp.100-105
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    • 2020
  • Northern pipevine (Aristolochia contorta) commonly inhabits marginal areas between waterside and terrestrial vegetation. In particular, A. contorta is ecologically important in the marginal areas as a food plant of dragon swallowtail butterfly (Sericinus montela), which is designated as vulnerable species in the Republic of Korea. For long-term sustainability of the plant population, assessment of the genetic diversity of exist populations should be conducted. Genomic DNA of A. contorta leaf samples were extracted from four populations where the vigorous growth were observed in the South Korea. Intra-population genetic diversity and inter-population genetic distance were assessed using randomly amplified polymorphic DNA (RAPD) with five polymorphic random primers. Overall genetic diversity was lower, compared to other wetland species (h: 0.0607 ~ 0.1401; I: 0.0819 ~ 0.1759), while GP showed the highest intra-population genetic diversity. Despite of the geographical distance, GP showed the larger genetic distance from other populations. This result seemed to be caused by the fragmented habitat and lower sexual reproduction of A. controta. Mixture of the different source populations and construction of the proper environmental condition such as shade and physical support for sexual reproduction should be considered for conservation of A. contorta population.

Reevaluation of the Change of Leuconostoc Species and Lactobacillus plantarum by PCR During Kimchi Fermentation

  • Choi, Jae-Yeon;Kim, Min-Kyun;Lee, Jong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.166-171
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    • 2002
  • The genus Leuconostoc is generally recognized as a favorable microorganism associated with a good taste of Kimchi and Lactobacillus plantarum is responsible for the overripening and acidification of Kimchi. A rapid and reliable PCR-based method to monitor the change of these lactic acid bacterial populations during Kimchi fermentation was attempted. A Leuconostoc-specific primer set was chosen from the conserved sequences of 16S rRNA genes among Leuconostoc species. The Lb. plantarum-specific primer set was the internal segments of a Lb. plantarum-specific probe which was isolated after randomly amplified polymorphic DNA (RAPD) analysis and tested for identification. The specificity of this protocol was examined in DNA samples isolated from a single strain. In agarose gel, as little as 10 pg of template DNA could be used to visualize the PCR products, and quantitative determination was possible at the levels of 10 pg to 100 ng template DNA. For the semi-quantitative determination of microbial changes during Kimchi fermentation, total DNAs from the 2 h-cultured microflora of Kimchi were extracted for 16 days and equal amounts of DNA templates were used for PCR. The intensities of DNA bands obtained from PCR using Leuconostoc-specific and Lb. plantarum-specific primer sets marked a dramatic contrast at the 1 ng and 100 ng template DNA levels during Kimchi fermentation, respectively. As the fermentation proceeded, the intensity of the band for Leuconostoc species increased sharply until the 5th day and the levels was maintained until the 11 th day. The sharp increase for Lb. plantarum occurred after 11 days with the decrease of Leuconostoc species. The results of this study indicate that Leuconostoc species were the major microorganisms at the beginning of Kimchi fermentation and reach their highest population during the optimum ripening period of Kimchi.

Characteristics of Dioscorea alata L. Introduced from Tropical and Subtropical Regions (도입 마(Dioscorea alata L.)의 특성 분석)

  • Chang, K.J.;Yoo, K.O.;Park, C.H.;Park, J.I.;Hong, K.H.;Park, J.H.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.3 no.1
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    • pp.48-69
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    • 2001
  • A lot of clones of the genus Dioscorea have been introduced from some tropical and subtropical regions since 1997. In 33 clones of water yams (Dioscorea alata L.), some morphological characteristics were investigated at the field. Variation ranges of the total weight and tuber number per stump were within the ranges from 90 to 2,147 g with an average of 610 g ; and 1.3-4.7 with an average of 2.8, respectively. The color tones observed on the tuber-flesh were sorted into 3 color-categories, i.e., white, pale brown and pale purple, and those on leaves were sorted into 3 color-categories, i.e., green, heavy green and purplish green. Intraspecific genetic relationship of 19 variation types of the Yam classified by their external morphological characteristics such as leaf and tuber shape was assessed by DNA using random and specific primers. Twenty two out of 113 primers (100 random[10-mer] primers, two 15 mer [M13 core sequence, and (GGAT)4 sequence]) had been used in PCR-amplification. Only 12 primers, however, were successful in DNA amplification in all of the analyzed plants, resulting in 93 randomly and specifically amplified DNA fragments. The analyzed taxa showed very high polymorphisms(69 bands, 71.0%), allowing individual taxon to be identified based on DNA fingerprinting. Monomorphic bands among total amplified DNA bands of each primer was low under the 50%. Similarity indices between accessions were computed from PCR(polymerase chain reaction) data, and genetic relationships among intraspecific variations were closely related at the levels ranging from 0.66 to 0.90.

The Relative Identification of C. officinale and L. chuanxiong by PCR-Mediated Fingerprinting (천궁류(川芎類) 한약재의 유전자 감식 연구)

  • Choi, Ho-Young;Kim, Dong-Wook;Kim, Dong-Eun;Suh, Young-Bae;Ham, In-Hye
    • The Korea Journal of Herbology
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    • v.20 no.4
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    • pp.151-161
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    • 2005
  • Objectives : Our research purpose is to establish the standard identification analysis on C. officinale and L. chuanxiong in Korea and China by PCR-mediated fingerprinting. Methods : The Restriction Fragment Length Polymorphism (RFLP) and Randomly Amplified Polymorphic DNA (RAPD) method was used on Internal Transcribed Spacer (ITS) regions and rbcL regions to compare and discriminate genes extracted from crude drugs as C. officinale and L. chuanxiong in Korea and China. Results : L. chuanxiong Korea and China have very similar polymorphism, whereas L. chuanxiong in Korea and C. officinale have very different polymorphism in RFLP. And restriction enzymes AluI and SacI forms the specific fragment band only in C. officinale, they can be used as RFLP marker on ITS regions to discriminate among the species. Conclusions : The results could be applied in discriminating crude drugs among C. officinale and L. chuanxiong in Korea and China. Also they could be used in controlling drug quality, preserving medicinal plants, and improving plant description.

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