• Title/Summary/Keyword: Gene Identification

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Isolation and Identification of Burkholderia pyrrocinia CH-67 to Control Tomato Leaf Mold and Damping-off on Crisphead Lettuce and Tomato

  • Lee, Kwang-Youll;Kong, Hyun-Gi;Choi, Ki-Hyuck;Lee, Seon-Woo;Moon, Byung-Ju
    • The Plant Pathology Journal
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    • v.27 no.1
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    • pp.59-67
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    • 2011
  • A bacterial strain CH-67 which exhibits antagonism towards several plant pathogenic fungi such as Botrytis cinerea, Fulvia fulva, Rhizoctonia solani, Sclerotinia sclerotiorum, Colletotrichum sp. and Phytophthora sp. was isolated from forest soil by a chitin-baiting method. This strain was identified as Burkholderia cepacia complex (Bcc) and belonging to genomovar IX (Burkholderia pyrrocinia) by colony morphology, biochemical traits and molecular method like 16S rRNA and recA gene analysis. This strain was used to develop a bio-fungicide for the control of tomato leaf mold caused by Fulvia fulva. Various formulations of B. pyrrocinia CH-67 were prepared using fermentation cultures of the bacterium in rice oil medium. The result of pot experiments led to selection of the wettable powder formulation CH67-C containing modified starch as the best formulation for the control of tomato leaf mold. CH67-C, at 100-fold dilution, showed a control value of 85% against tomato leaf mold. Its disease control efficacy was not significantly different from that of the chemical fungicide triflumidazole. B. pyrrocinia CH-67 was also effective in controlling damping-off caused by Rhizoctonia solani PY-1 in crisphead lettuce and tomato plants. CH67-C formulation was recognized as a cell-free formulation since B. pyrrocinia CH-67 was all lethal during formulation process. This study provides an effective biocontrol formulation of biofungicide using B. pyrrocinia CH-67 to control tomato leaf mold and damping-off crisphead lettuce and tomato.

Xanthomonas euvesicatoria Causes Bacterial Spot Disease on Pepper Plant in Korea

  • Kyeon, Min-Seong;Son, Soo-Hyeong;Noh, Young-Hee;Kim, Yong-Eon;Lee, Hyok-In;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.32 no.5
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    • pp.431-440
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    • 2016
  • In 2004, bacterial spot-causing xanthomonads (BSX) were reclassified into 4 species-Xanthomonas euvesicatoria, X. vesicatoria, X. perforans, and X. gardneri. Bacterial spot disease on pepper plant in Korea is known to be caused by both X. axonopodis pv. vesicatoria and X. vesicatoria. Here, we reidentified the pathogen causing bacterial spots on pepper plant based on the new classification. Accordingly, 72 pathogenic isolates were obtained from the lesions on pepper plants at 42 different locations. All isolates were negative for pectolytic activity. Five isolates were positive for amylolytic activity. All of the Korean pepper isolates had a 32 kDa-protein unique to X. euvesicatoria and had the same band pattern of the rpoB gene as that of X. euvesicatoria and X. perforans as indicated by PCR-restriction fragment length polymorphism analysis. A phylogenetic tree of 16S rDNA sequences showed that all of the Korean pepper plant isolates fit into the same group as did all the reference strains of X. euvesicatoria and X. perforans. A phylogenetic tree of the nucleotide sequences of 3 housekeeping genes-gapA, gyrB, and lepA showed that all of the Korean pepper plant isolates fit into the same group as did all of the references strains of X. euvesicatoria. Based on the phenotypic and genotypic characteristics, we identified the pathogen as X. euvesicatoria. Neither X. vesicatoria, the known pathogen of pepper bacterial spot, nor X. perforans, the known pathogen of tomato plant, was isolated. Thus, we suggest that the pathogen causing bacterial spot disease of pepper plants in Korea is X. euvesicatoria.

Identification of Upregulated APOA1 Protein of Chicken Liver in Pullorum Disease (추백리가 감염된 닭의 간에서 발현이 증가하는 APOA1 단백질의 확인)

  • Jung K. C.;Lee Y. J.;Yu S. L.;Lee J. H.;Jang B. K.;Koo Y. B.;So H. K.;Choi K. D.
    • Korean Journal of Poultry Science
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    • v.32 no.1
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    • pp.23-27
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    • 2005
  • The aim of this study was to investigate differentially expressed proteins between normal chicken liver and chicken liver inffeted by Salmonella pullorum. 2-dimensional electrophoresis (2DE) and mass spectrometry (MS) were used to identify the proteins. More than 300 protein spots were detected on silver stained 2DE gels using pH 3$\~$10 gradients. The most outstanding protein spot was further analyzed by MALDI-TOF MS and protein database using the Mascot search engine. The protein was finally identified as APOAI (Apolipoprotein AI). Based on the known function of the APOAI, this gene acts protective action against the accumulation of platelet thrombin at the site of vascular damage for the pullorum disease. Therefore APOAI protein, identified in this study, can be a valuable biomarker in relation to the pullorum disease in chicken.

Isolation and identification of Vibrio harveyi from chub mackerel (Scomber japonicus)

  • Lee, Young-Ran;Jun, Jin-Woo;Giri, Sib Sankar;Kim, Hyoun-Joong;Yun, Sae-Kil;Chi, Cheng;Kim, Sang-Guen;Koh, Jeong-Rack;Jung, Ji-Yun;Park, Se Chang
    • Korean Journal of Veterinary Research
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    • v.56 no.1
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    • pp.45-46
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    • 2016
  • For several days, there was a series of mortalities of chub mackerel (Scomber japonicus) that were reared for public exhibition in a private aquarium in Seoul, Korea. As part of the diagnosis of the dead fish, a bacterial isolate from the kidney was cultured, identified, and confirmed to be Vibrio (V.) harveyi using Vitek System 2 and 16S rRNA gene sequencing. Phylogenetic analysis was also performed by the neighbor-joining method. As a result, the V. harveyi isolated from chub mackerels of a private aquarium in Korea, called as SNUVh-LW1, was clustered in the same group with V. harveyi ATCC33843.

Cyanobacterial Diversity Analysis Using cpcBA-Intergenic Spacer Region (cpcBA-Intergenic Spacer Region을 이용한 Cyanobacteria의 다양성 분석)

  • Choi Gang-Guk;Park Yong-Ha;Ahn Chi-Yong;Bae Myoung-Sook;Oh Hee-Mock
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.287-292
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    • 2005
  • The cyanobacterial diversity was analyzed by restriction fragment length polymorphism (RFLP) of PCR-amplified rpcBA-Intergenic Spacer (IGS) genes and cpcBA-IGS gene sequencing with a sample collected at Chuso-ri in Daechung Reservoir on March 15, 2005, The Shannon-Weiner diversity index was 0.65, indicating that the cyanobacterial community structure was simple. PCR-RFLP profiles obtained were Phormidium spp. (58 clones), Anabaena spp. (14 clones), Microcystis spp. (4 clones), Spirulina sp. (1 clone) and uncultured cyanobacteria (2 clones). The PCR-RFLP of cpcBA-IGS revealed that Phormidium spp. and Anabaena spp. dominated in the invested sample. As a consequence, it seems that the analysis of functional genes such as cpcBA-IGS can be used for the species identification and community analysis of cyanobacteria.

Identification and Cloning of jipA Encoding a Polypeptide That Interacts with a Homolog of Yeast Rad6, UVSJ in Aspergillus nidulans

  • Cho, Jae-Han;Yun, Seok-Soong;Jang, Young-Kug;Cha, Mee-Jeong;Kwon, Nak-Jung;Chae, Suhn-Kee
    • Journal of Microbiology
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    • v.41 no.1
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    • pp.46-51
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    • 2003
  • RAD6 in yeast mediates postreplication DNA repair and is responsible for DNA-damage induced mutations. RAD6 encodes ubiquitin-conjugating enzyme that is well conserved among eukaryotic organisms. However, the molecular targets and consequences of their ubiquitination by Rad6 have remained elusive. In Aspergillus nidulans, a RAD6 homolog has been isolated and shown to be an allele of uvs). We screened a CDNA library to isolate UVSJ-interacting proteins by the yeast two-hybrid system. JIPA was identified as an interactor of UVSJ. Their interaction was confirmed in vitro by a GST-pull down assay. JIPA was also able to interact with mutant UVSJ proteins, UVSJl and the active site cysteine mutant UVSJ-C88A. The N- and the C-terminal regions of UVSJ required for the interaction with UVSH, a RAD18 homolog of yeast which physically interacts with Rad6, were not necessary for the JIPA and UVSJ interactions. About 1.4 kb jipA transcript was detected in Northern analysis and its amount was not significantly increased in response to DNA-damaging agents. A genomic DNA clone of the jipA gene was isolated from a chromosome I specific genomic library by PCR-sib selection. Sequence determination of genomic and cDNA of jipA revealed an ORF of 893 bp interrupted by 2 introns, encoding a putative polypeptide of 262 amino acids. JIPA has 33% amino acid sequence identity to TIP41 of Saccharomyces cerevisiae which negatively regulates the TOR signaling pathway.

Identification of salt and drought inducible glutathione S-transferase genes of hybrid poplar

  • Kwon, Soon-Ho;Kwon, Hye-Kyoung;Kim, Wook;Noh, Eun Woon;Kwon, Mi;Choi, Young Im
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.26-32
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    • 2014
  • Recent genome annotation revealed that Populus trichocarpa contains 81 glutathione S-transferase (GST) genes. GST genes play important and varying roles in plants, including conferring tolerance to various abiotic stresses. Little information is available on the relationship - if any - between drought/salt stresses and GSTs in woody plants. In this study, we screened the PatgGST genes in hybrid poplar (Populus alba ${\times}$ Populus tremula var. glandulosa) that were predicted to confer drought tolerance based on our expression analysis of all members of the poplar GST superfamily following exposure to salt (NaCl) and drought (PEG) stresses, respectively. Exposure to the salt stress resulted in the induction of eight PatgGST genes and down-regulation of one PatgGST gene, and the level of induction/repression was different in leaf and stem tissues. In contrast, 16 PatgGST genes were induced following exposure to the drought (PEG) stress, and two were down-regulated. Taken together, we identified seven PatgGSTs (PatgGSTU15, PatgGSTU18, PatgGSTU22, PatgGSTU27, PatgGSTU46, PatgGSTU51 and PatgGSTU52) as putative drought tolerance genes based on their induction by both salt and drought stresses.

Characterization of Homologous Defective Interfering RNA during Persistent Infection of Vero Cells with Japanese Encephalitis Virus

  • Yoon, Sung Wook;Lee, Sang-Yong;Won, Sung-Yong;Park, Sun-Hee;Park, Soo-Young;Jeong, Yong Seok
    • Molecules and Cells
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    • v.21 no.1
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    • pp.112-120
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    • 2006
  • It has been suggested that defective interfering (DI) RNA contributes to the persistence of Japanese encephalitis virus (JEV). In this study, we characterized molecular and biological aspects of the DI RNA and its relation to viral persistence. We identified a homologous DI virus intimately associated with JEV persistence in Vero cells. The production of DI RNA during undiluted serial passages of JEV coincided with the appearance of cells refractory to acute infection with JEV. We also established a Vero cell clone with a persistent JEV infection in which the DI RNA coreplicated efficiently at the expense of helper virus. The infectious virus yield of the clone fluctuated during its growth depending upon the amount of DI RNA accumulated in the previous replication cycle. Identification of the corresponding negative-sense RNA of the DI RNA indicated that the DI RNA functioned as a replication unit. Most of the DI RNA molecules retained their open reading frames despite a large deletion, encompassing most of the prM, the entire E, and the 5' half of the NS1 gene. Taken together, these observations suggest that the generation of homologous DI RNA during successive JEV acute infections in Vero cells probably participates actively in persistent JEV infection.

Molecular identification and expression analysis of bactericidal permeability-increasing protein/ LPS-binding protein (BPI/LBP) from Black rockfish Sebastes schlegeli

  • Kwon, Mun-Gyeong;Kim, Ju-Won;Park, Myoung-Ae;Hwang, Jee-Youn;Park, Hyung-Jun;Baeck, Gun-Wook;Park, Chan-Il
    • Journal of fish pathology
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    • v.23 no.3
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    • pp.323-334
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    • 2010
  • Bactericidal/permeability-increasing protein (BPI) and lipopolysaccharide-binding protein (LBP) are important components of the mammalian innate defence system against Gram-negative infections. The BPI/LBP cDNA was identified from the black rockfish ConA/PMA or LPS stimulated leukocyte cDNA library. The full-length BR-BPI/LBP cDNA was 2118 bp long and contained an open reading frame (ORF) of 1422 bp that encoded 473 amino-acid residues. The 5' UTR had a length of 57 bp, and the 3' UTR 639 bp. The molecular weight and theoretical isoelectric point (pI) values were calculated 51.4 kDa and 9.72, respectively. Compared with other known BPI or BPI/LBP peptide sequences, the most conserved regions of the black rockfish BPI/LBP peptide were found to be the BPI1 N-terminal, BPI2 C-terminal domains and a LPS binding domain. Phylogenetic analysis based on the deduced amino acid sequence revealed a homologous relationship between the BPI/LBP sequence of black rockfish and that of other teleosts. The black rockfish BPI/LBP gene was predominantly expressed in the PBLs, head kidney, trunk kidney and spleen. The expression of the black rockfish BPI/LBP molecule was induced in the peripheral blood leukocytes (PBLs) from 1 to 24 h following LPS stimulation, with a peak at 12 h post-stimulation.

Physicochemical and Microbiological Characterization of Protected Designation of Origin Ezine Cheese: Assessment of Non-starter Lactic Acid Bacterial Diversity with Antimicrobial Activity

  • Uymaz, Basar;Akcelik, Nefise;Yuksel, Zerrin
    • Food Science of Animal Resources
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    • v.39 no.5
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    • pp.804-819
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    • 2019
  • Ezine cheese is a non-starter and long-ripened cheese produced in the Mount of Ida region of Canakkale, Turkey, with a protected designation of origin status. Non-starter lactic acid bacteria (NSLAB) have a substantial effect on the quality and final sensorial characteristics of long-ripened cheeses. The dominance of NSLAB can be attributed to their high tolerance to the hostile environment in cheese during ripening relative to many other microbial groups and to its ability to inhibit undesired microorganisms. These qualities promote the microbiological stability of long-ripened cheeses. In this study, 144 samples were collected from three dairies during the ripening period of Ezine cheese. Physicochemical composition and NSLAB identification analyses were performed using both conventional and molecular methods. According to the results of a 16S rRNA gene sequence analysis, 13 different species belonging to seven genera were identified. Enterococcus faecium (38.42%) and E. faecalis (18.94%) were dominant species during the cheese manufacturing process, surviving 12 months of ripening together with Lactobacillus paracasei (13.68%) and Lb. plantarum (11.05%). The results indicate that NSLAB contributes to the microbiological stability of Ezine cheese over 12 months of ripening. The isolation of NSLAB with antimicrobial activity, potential bacteriocin producers, yielded defined collections of natural NSLAB isolates from Ezine cheese that can be used to generate specific starter cultures for the production of Ezine cheese (PDO).