• Title/Summary/Keyword: DNA fingerprinting

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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.

Genetic and Phenotypic Diversity of Fenitrothion-Degrading Bacteria Isolated from Soils

  • Kim, Kyung-Duk;Ahn, Jae-Hyung;Kim, Tae-Sung;Park, Seong-Chan;Seong, Chi-Nam;Song, Hong-Gyu;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.19 no.2
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    • pp.113-120
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    • 2009
  • Twenty-seven fenitrothion-degrading bacteria were isolated from different soils, and their genetic and phenotypic characteristics were investigated. Analysis of the 16S rDNA sequence showed that the isolates were related to members of the genera Burkholderia, Pseudomonas, Sphingomonas, Cupriavidus, Corynebacterium, and Arthrobacter. Among the 27 isolates, 12 different chromosomal DNA fingerprinting patterns were obtained by polymerase chain reaction(PCR) amplification of repetitive extra genic palindromic(REP) sequences. The isolates were able to utilize fenitrothion as a sole source of carbon and energy, producing 3-methyl-4-nitrophenol as the intermediate metabolite during the complete degradation of fenitrothion. Twenty-two of 27 isolates were able to degrade parathion, methyl-parathion, and p-nitrophenol but only strain BS2 could degrade EPN(O-ethyl-O-p-nitrophenyl phenylphosphorothioate) as a sole source of carbon and energy for growth. Eighteen of the 27 isolates had plasmids. When analyzed with PCR amplification and dot-blotting hybridization using various specific primers targeted to the organophosphorus pesticide hydrolase genes of the previously reported isolates, none of the isolates showed positive signals, suggesting that the corresponding genes of our isolates had no significant sequence homology with those of the previously isolated organophosphate pesticide-degrading bacteria.

Characterization of Hemolytic Aeromonas sp. MH-8 Responding to the Exposure of Green Tea Catechin, EGCG (녹차 카테킨 EGCG의 노출에 따른 식중독 세균인 용혈성 Aeromonas sp. MH-8의 특성조사)

  • Kim, Dong-Min;Oh, Kye-Heon
    • KSBB Journal
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    • v.31 no.4
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    • pp.228-236
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    • 2016
  • The aim of this study was to characterize the hemolytic Aeromonas sp. MH-8 exposed to green tea catechin, epigallocatechin gallate (EGCG). Initially, the hemolytic Aeromonas sp. MH-8 was enriched and isolated from stale fish. Bactericidal effects of MH-8 exposed to EGCG ranging from 1 mg/mL to 4 mg/mL were monitored, and complete bactericidal effects were achieved within 3 h at 3 mg/mL and higher concentrations. SDS-PAGE with silver staining revealed that the amount of lipopolysaccharides increased or decreased in the strain MH-8 treated to different concentrations and exposing periods of EGCG in exponentially growing cultures. The stress shock proteins (70-kDa DnaK and 60-kDa GroEL), which might contribute to enhancing the cellular resistance to the cytotoxic effect of EGCG, were induced at different concentrations of EGCG exposed to cell culture of MH-8. Scanning electron microscopic analysis demonstrated the presence of irregular rod shapes with umbilicated surfaces for cells treated with EGCG. 2-DE of soluble protein fractions from MH-8 cultures showed 18 protein spots changed by EGCG exposure. These proteins involved in chaperons (e.g., DnaK, GroEL and trigger factor), enterotoxins (e.g., aerolysin and phospholipase C precursor), LPS synthesis (e.g., LPS biosynthesis protein and outer membrane protein A precursor), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. In consequence, EGCG was found to have substantial antibacterial effects against food-poisoning causing bacterium, hemolytic Aeromonas sp. MH-8. Also the results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on Aeromonas sp. MH-8.

Development of SNP marker set for discriminating among Korean rice varieties and imported rice in Korea

  • Park, Seul-Gi;Lee, Hyo-Jeong;Lee, Keon-Mi;Baek, Man-Kee;Park, Hyun-Su;Shin, Woon-Chul;Nam, Jeong-Kwon;Kim, Choon-Song;Kim, Bo-Kyeong;Cho, Young-Chan
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.154-154
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    • 2017
  • In accordance with the opening of the Korean rice market, this study was focused on establishment of database for discriminating the Korean rice varieties and imported brand rices using DNA markers. In this study, the SNP markers were developed using single nucleotide polymorphisms between the reference sequences of japonica and them of 40 brand rices which collected in Australia, China, Thailand, United States and Vietnam. The developed SNP markers were screened to a total of 360 rices including 320 Korean rice varieties and 40 imported brand rices. We selected polymorphic markers among Korean bred rive varieties and imported brand rices. The selected markers were classified into 3 grades. The markers of A grade produced DNA band in 360 rices of 30~40%, B grades produced in 40~60%, and C grades produced bands over 60% rices. First, we tried to set-up the discriminating system using the minimum SNP markers of A grade. Especially, a set of sixteen SNP markers could identify among Korean bred rice varieties and imported brand rices. Additionally, some SNP markers like NSb for Pib gene, JJ80-T for Pi5 and YL155/YL87 for Pita which linked to resistance genes to blast were used to fingerprinting system. These markers were set-up as multiplex set for enhancing the identification efficiency among rice varieties. Finally, the selected SNP markers would be used to the fluidigm assay to construct the database for elaborate discrimination of rice varieties.

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Identification of Bacteriocin-Producing Lactobacillus paraplantarum First Isolated from Kimchi

  • LEE JONG HOON;KIM MUIN;JEONG DO WON;KIM MIN JUNG;KIM JEONG HWAN;CHANG HAE CHOON;CHUNG DAE KYUN;KIM HAE YEONG;KIM KYOUNG HEON;LEE HYONG JOO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.428-433
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    • 2005
  • A bacteriocin-producing lactic acid bacterium with inhibitory activity against the growth of Lactobacillus plantarum was isolated from kimchi, a traditional Korean fermented vegetable. For the identification of the isolate, its 16S rDNA was sequenced. As a result, the sequence showed $99\%$ homology with those from Lactobacillus paraplantarum, Lb. plantarum, and Lactobacillus pentosus. For further identification of the isolate, the sequence of its 16S/23S rDNA spacer region was determined, and the sequence matched perfectly with that of Lb. paraplantarum. SDS­PAGE fingerprinting of whole-cell proteins of the isolate was almost identical with that of Lb. paraplantarum. The isolation and identification of Lb. paraplantarum suggest that Lb. paraplantarum is one of the lactic acid bacteria involved in kimchi fermentation.

Genetic Diversity of Paecilomyces japonica and Cordypces militaris Strains by URP-PCR Fingerprinting (URP-PCR핵산지문에 의한 눈꽃동충하초 (Paecilomyces japonica.)와 번데기동충하초(Cordypces militaris) 유전적 다양성분석)

  • Kim, Jong-Kun;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.39 no.3
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    • pp.180-184
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    • 2011
  • This study was carried out to identify the genetic characteristics among isolates of Paecilomyces spp.and Cordyceps spp. by URP-PCR analysis. Twenty URP (universal rice primer) primers of 20 mer which were designed from repetitive sequence of rice, were used for producing PCR DNA fingerprints of the mushrooms. Of them, 5 URP primers, URP2F, URP2R, URP9F, URP4R, and URP17R amplified genomic DNA of the mushrooms with polymorphic PCR patterns. On isolates of Cordyceps militaris, primers URP1F, URP2R, URP6R and URP17R produced PCR polymorphic bands of 4 types. Isolates of Cordypces sp. that are isolated from different area of Korea were identical to isolate of C. militaris, while other species of Cordypces were different to the PCR profiles. However, the URP primers did not identify the polymorphism of PCR profile on isolates of P. japonica.

Genetic Diversity and Phylogenetic Relationships among Microsporidian Isolates from the Indian Tasar Silkworm, Antheraea mylitta, as Revealed by RAPD Fingerprinting Technique

  • Hassan, Wazid;Nath, B. Surendra
    • International Journal of Industrial Entomology and Biomaterials
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    • v.29 no.2
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    • pp.169-178
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    • 2014
  • In this study, we investigated genetic diversity of 22 microsporidian isolates infecting tropical tasar silkworm, Antheraea mylitta collected from various geographical forest locations in the state of Jharkhand, India, using polymerase chain reaction (PCR)-based marker assay: random amplified polymorphic DNA (RAPD). A type species, NIK-1s_mys was used as control for comparison. The shape of mature microsporidians was found to be oval to elongate, measuring 3.80 to $5.10{\mu}m$ in length and 2.56 to $3.30{\mu}m$ in width. Of the 20 RAPD primers screened, 16 primers generated reproducible profiles with 298 polymorphic fragments displaying high degree of polymorphism (97%). A total of 14 RAPD primers produced 45 unique putative genetic markers, which were used to differentiate the microsporidians. Calculation of genetic distance coefficients based on dice coefficient method and clustering with un-weighted pair group method using arithmetic average (UPGMA) analysis was conducted to unravel the genetic diversity of microsporidians infecting tasar silkworm. The similarity coefficients varied from 0.059 to 0.980. UPGMA analysis generated a dendrogram with four microsporidian groups, which appear to be different from each other as well as from NIK-1s_mys. Two-dimensional distribution based on Euclidean distance matrix also revealed considerable variability among different microsporidians identified from the tasar silkworms. Clustering of few microsporidian isolates was in accordance with the geographic origin. The results indicate that the RAPD profiles and specific/unique genetic markers can be used for differentiating as well as to identify different microsporidians with considerable accuracy.

Analysis of intraspecific genetic diversity in Acidovorax citrulli causing bacterial fruit blotch on cucurbits in Korea

  • Song, Jeong Young;Oo, May Moe;Park, Su Yeon;Seo, Mun Won;Lee, Seong-Chan;Jeon, Nak Beom;Nam, Myeong Hyeon;Lee, Youn Su;Kim, Hong Gi;Oh, Sang-Keun
    • Korean Journal of Agricultural Science
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    • v.45 no.4
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    • pp.575-582
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    • 2018
  • Bacterial fruit blotch (BFB) caused by Acidovorax citrulli is a devastating disease found in many cucurbits cultivation fields. The genetic diversity for 29 strains of A. citrulli collected from various cucurbits in South Korea was determined by DNA fingerprinting with a pathogenicity test, multi locus analysis, Rep-PCR (repetitive sequence polymerase chain reaction), and URP (universal rice primers) PCR bands. Two distinct groups (Korean Clonal Complex, KCC1 and KCC2) in the population were identified based on group specific genetic variation in the multi locus phylogeny using six conserved loci and showed a very high similarity with DNA sequences for representative foreign groups [the group I (CC1-1 type) and the group II (CC2-5 type)] widely distributed worldwide, respectively. Additionally, in the case of phaC, a new genotype was found within each Korean group. The KCC1 was more heterogeneous compared to the KCC2. The KCC1 recovered mainly from melons and watermelons (ratio of 6 : 3) and 15 of the 20 KCC2 strains recovered from watermelons were dominant in the pathogen population. Accordingly, this study found that two distinct groups of differentiated A. citrulli exist in South Korea, genetically very similar to representative foreign groups, with a new genotype in each group resulting in their genetic diversity.

Thermophilic Biofiltration of Benzene and Toluene

  • Cho, Kyung-Suk;Yoo, Sun-Kyung;Ryu, Hee-Wook
    • Journal of Microbiology and Biotechnology
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    • v.17 no.12
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    • pp.1976-1982
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    • 2007
  • In the current studies, we characterized the degradation of a hot mixture of benzene and toluene (BT) gases by a thermophilic biofilter using polyurethane as a packing material and high-temperature compost as a microbial source. We also examined the effect of supplementing the biofilter with yeast extract (YE). We found that YE substantially enhanced microbial activity in the thermophilic biofilter. The degrading activity of the biofilter supplied with YE was stable during long-term operation (approximately 100 d) without accumulating excess biomass. The maximum elimination capacity ($1,650\;g{\cdot} m^{-3}{\cdot} h^{-1}$) in the biofilter supplemented with YE was 3.5 times higher than that in the biofilter without YE ($470\;g{\cdot} m^{-3}{\cdot} h^{-1}$). At similar retention times, the capacity to eliminate BT for the YE-supplemented biofilter was higher than for previously reported mesophilic biofilters. Thus, thermophilic biofiltration can be used to degrade hydrophobic compounds such as a BT mixture. Finally, 168 rDNA polymerase chain reaction-DGGE (PCR-DGGE) fingerprinting revealed that the thermophilic bacteria in the biofilter included Rubrobacter sp. and Mycobacterium sp.

Pathogenic and Molecular Characteristics of Agrobacterium vitis strains isolated from Grapevine in Korea

  • Kim, J.G.;Kim, S.H.;Choi, J.E.;Lee, Y.K.;Kang, H.W.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.120.2-120
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    • 2003
  • Agrobacterium vitis is a causal agent of crown-gall disease on grapevine. In Korea, grapevine variety (GeoBong) have severely been infected by the bacteria since stems of the variety were buried in soil for overwintering. Infection ratio over 70-80% was observed on 7 years old GeoBong grapevine in Ansung and Cheonan. PCR specific primers for A. vitis strains were designed using nucleotide sequences of vir A gene in Ti-Plasmid, pheA gene in chromosomal DNA and a URP-PCR polymorphic band. Three hundred bacterial strains were isolated from the different 80 galls formed on GeoBong grapevine in Cheonan and Ansung of Korea and were screened to identify A. vitis using the three specific PCR primers for Agrobacterium vitis. Twenty-four bacterial strains that are detected by the primers were further confirmed by pathogenicity and biochemical methods. To investigate the genomic diversity of the bacterial strains, twenty primers of 20 mer referred to universal rice primers (URP) were applied for PCR fingerprinting, Of them, URP2R and URP2F primers could effectively be used to detect polymorphism within the bacterial strains.

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