• Mycobiology
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• v.42 no.2
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• pp.174-180
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• 2014

We analyzed the genetic diversity of Cylindrocarpon destructans isolates obtained from Korean ginseng (i.e., Panax ginseng) roots by performing virulence tests and nuclear ribosomal gene internal transcribed spacer (ITS) and mitochondrial small subunit (mt SSU) rDNA sequence analysis. The phylogenetic relationship analysis performed using ITS DNA sequences and isolates from other hosts helped confirm that all the Korean C. destructans isolates belonged to Nectria/Neonectria radicicola complex. The results of in vivo and ex vivo virulence tests showed that the C. destructans isolates could be divided into two groups according to their distinctive difference in virulence and the genetic diversity. The highly virulent Korean isolates in pathogenicity group II (PG II), together with foreign isolates from P. ginseng and P. quinquefolius, formed a single group. The weakly virulent isolates in pathogenicity group I, together with the foreign isolates from other host plants, formed another group and exhibited a greater genetic diversity than the isolates of PG II, as confirmed by the mt SSU rDNA sequence analysis. In addition, as the weakly virulent Korean isolates were genetically very similar to the foreign isolates from other hosts, they were likely to originate from hosts other than the ginseng plants.

• The Plant Pathology Journal
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• v.33 no.3
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• pp.249-263
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• 2017

This study aims to examine the potential reasons for the current prevalence of the fusarium wilt in the oriental melon. Twenty-seven Fusarium isolates obtained from oriental melon greenhouses in 2010-2011 were identified morphologically and by analysis of elongation factor-1 alpha gene (EF-$1{\alpha}$) and internal transcribed spacer (ITS) rDNA sequences as 6 Fusarium species (8 isolates of F. oxysporum, 8 F. commune, 5 F. proliferatum, 3 F. equiseti, 2 F. delphinoides, and 1 F. andiyazi), which were classified as same into 6 EF-$1{\alpha}$ sequence-based phylogenetic clades. Pathogenicity of the Fusarium isolates on the oriental melon was highest in F. proliferatum, next in F. oxysporum and F. andiyazi, and lowest in the other Fusarium species tested, suggesting F. proliferatum and F. oxysporum were major pathogens of the oriental melon, inducing stem rots and vascular wilts, respectively. Oriental melon and watermelon were more susceptible to F. oxysporum than shintosa and cucumber; and cucumber was most, oriental melon and watermelon, medially, and shintosa was least susceptible to F. proliferatum, whose virulence varied among and within their phylogenetic subclades. Severe root-knot galls were formed on all the crops infected with Meloidogyne incognita; however, little indication of vascular wilts or stem and/or root rots was shown by the nematode infection. These results suggest the current fungal disease in the oriental melon may be rarely due to virulence changes of the fusarium wilt pathogen and the direct cause of the severe root-knot nematode infection, but may be potentially from other Fusarium pathogen infection that produces seemingly wilting caused by severe stem rotting.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.5
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• pp.582-588
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• 2016

Seventy-nine Vibrio parahaemolyticus isolates from surface seawater from Gomso Bay, west coast of Korea, were analyzed for the presence of virulence genes and their susceptibility to 30 different antimicrobials. All 79 isolates were examined for the presence of two virulence genes (tdh or trh) using polymerase chain reaction (PCR); however, no isolates possessed either the tdh or trh gene. According to a disk diffusion susceptibility test, all of the strains studied were resistant to oxacillin, penicillin, and vancomycin, followed by ticarcillin (97.5%), ampicillin (96.2%), clindamycin (86.1%), erythromycin (10.1%), streptomycin (7.6%), cefoxitin (6.3%), amikacin (2.5%), and cephalothin (2.5%). However, all of the strains were susceptible to 19 other antimicrobials including cefepime, cefotaxime, chloramphenicol, gentamycin, nalidixic acid, sulfamethoxazole/trimethoprim, and trimethoprim. All 79 isolates (100%) were resistant to four or more classes of antimicrobials, and two strains exhibited resistance to eight antimicrobial agents. The average minimum inhibitory concentrations (MICs) for V. parahaemolyticus for ampicillin, penicillin, ticarcillin, and vacomycin were 946.5, 1,305.9, 1,032.3, and 45.0 µg/mL, respectively.

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.59-62
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• 2002

The role of a TolC homologue in the virulence of Vibrio vulnificus, a marine bacterium causing serious wound infection and fulminant septicemia in persons with underlying conditions, has been studied. TolC, an outer membrane protein, has been implicated in a variety of bacterial functions including export of diverse molecules ranging from large proteins to antibiotics. A homologue of the tolC gene of V. cholerae, which has been shown to be required for bile resistance, cytotoxicity and colonization of this organism, was identified in the partially determined genome sequence of V. vulnificus. To determine the role of TolC in the virulence of V. vulnificus, a TolC-deficient (TD) mutant was isolated by in vivo allelic exchange. Compared with the parent strain, the TD mutant was more sensitive to bile, and much less virulent in mice challenged subcutaneously. This mutant was noncytotoxic to the HEp-2 cells, but its metalloprotease and cytolysin activities in the culture supernatant were comparable to the parent strain. In addition, the resistance of the TD mutant to human serum bactericidal activity as well as its growth in either human or murine blood was not affected. Collectively, our data suggest that TolC may be involved in colonization and/or spread of V. vulnificus to the blood stream, probably by secreting a cytotoxin other than the cytolysin.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.52 no.6
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• pp.587-595
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• 2019

Eighty-three Vibrio parahaemolyticus isolates from surface seawater in Gomso Bay on the west coast of Korea, and commercial fisheries from Gunsan fisheries center were analyzed for the presence of virulence genes and susceptibility to 30 different antimicrobials. All 83 isolates were examined for the presence of two virulence genes (tdh or trh) using polymerase chain reaction; however, neither gene was found in any of the isolates. A disk diffusion susceptibility test, showed that all of the strains studied were resistant to clindamycin, oxacillin, ticarcillin, and vancomycin, and also revealed varying levels of resistance to ampicillin (98.8%), penicillin G (95.2%), streptomycin (20.5%), cefoxitin (14.5%), amikacin (6.0%), cephalothin (4.8%), and erythromycin (3.6%). However, all of the strains were susceptible to 19 other antimicrobial agents, including cefepime, cefotaxime, chloramphenicol, gentamycin, nalidixic acid, sulfamethoxazole/trimethoprim, and trimethoprim. All 83 isolates (100%) were resistant to five or more classes of antimicrobials, and two strains exhibited resistance to ten antimicrobial agents. The average minimum inhibitory concentrations against V. parahaemolyticus of clindamycin, oxacillin, ticarcillin, and vancomycin were 55.9, 98.3, 499.3, and 44.3 ㎍/mL, respectively. These results provide new insight into the necessity for seawater sanitation in Gomso Bay and commercial fisheries, and provide evidence to help reduce the risk of contamination by antimicrobial-resistant bacteria.

• The Plant Pathology Journal
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• v.30 no.3
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• pp.304-309
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• 2014

The rpf genes and $colS_{XOO1207}/colR_{XOO1208}$ were known to require for virulence of Xanthomonas oryzae pv. oryzae (Xoo). In Xoo KACC10331 genome, two more colS/colR genes, $colS_{XOO3534}$ (raxH)/$colR_{XOO3535}$ (raxR) and $colS_{XOO3762}/colR_{XOO3763}$ were annotated. The $colS_{XOO3534}/colR_{XOO3535}$ were known to control AvrXa21 activity and functions of $colS_{XOO3762}/colR_{XOO3763}$ were unknown in Xoo. To characterize the relationship between rpf and colS/colR genes, expression of colS/colR genes in Rpf mutants of Xoo were analyzed with quantitative reverse transcription PCR (qRT-PCR). Expressions of all three colS/colR genes increased in the rpfF mutant in which DSF synthesis is defective. Expression of $colS_{XOO1207}/col-R_{XOO1208}$, $colS_{XOO3534}/colR_{XOO3535}$ and $colS_{XOO3762}/colR_{XOO3763}$ increased 2, 2-7, 3-13 folds respectively. Expression of $colS_{XOO3534}$ and $colS_{XOO3762}$ also increased 2-4 folds in the rpfG mutant in which the signal from DSF is no longer transferred to down-stream. Expression of the other colS/colR genes was not significantly changed in the rpfG mutant compared to the wild type. Since RpfF and RpfG are responsible for DSF synthesis and signal transfer from DSF to down-stream to regulate virulence gene expression, these results suggest that the DSF and DSF-mediated signal regulate negatively three colS/colR genes in Xoo.

• Korean Journal of Veterinary Research
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• v.54 no.1
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• pp.39-48
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• 2014

The prevalence of thermophilic Campylobacter (C.) spp. in stray, breeding, and household dogs was 25.2, 12.0, and 8.8%, respectively. C. jejuni and C. upsaliensis were the predominant Campylobacter spp. from household dogs. cdtA, cdtB, and cdtC were detected by PCR in all isolates. Despite the high cytolethal distending toxin (CDT) gene prevalence, only 26 (31%) C. jejuni strains and one (15.3%) C. coli strain showed evidence of CDT production in HEp-2 cell cytotoxicity assays. Virulence-associated genes detected in the C. jejuni and C. coli isolates were cadF, dnaJ, flaA, racR, ciaB, iamA, pldA, virB11, ceuE, and docC. cadF, dnaJ, flaA, and ceuE were found in all C. jejuni and C. coli isolates. When detecting Guillain-Barr$\acute{e}$ syndrome-associated genes (galE, cgtB, and wlaN), galE was identified in all isolates. However, cgtB and wlaN were more prevalent in C. jejuni isolates from humans than those from dogs. Adherence and invasion abilities of the C. jejuni and C. coli strains were tested in INT-407 cells. A considerable correlation (adjusted $R^2$= 0.678) existed between adherence and invasion activities of the Campylobacter spp. isolates.

• The Plant Pathology Journal
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• v.35 no.2
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• pp.125-136
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• 2019

Vitis vinifera is very susceptible to downy mildew (Plasmopara viticola). A number of authors have suggested different genetic populations of this fungus exist in Europe, each showing a different degree of virulence. Work performed to date indicates this diversity to be the result of different factors. In areas where gene flow is greater and recombination more frequent, the diversity of P. viticola appears to be wider. In vineyards isolated by geographic barriers, a race may become dominant and produce clonal epidemics driven by asexual reproduction. The aim of the present work was to identify the conditions that influence the genetic diversity of P. viticola populations in the vineyards of northwestern Spain, where the climatic conditions for the growth of this fungus are very good. Vineyards situated in a closed, narrow valley of the interior, in more open valleys, and on the coast were sampled and the populations of P. viticola detected were differentiated at the molecular level through the examination of microsatellite markers. The populations of P. viticola represented in primary and secondary infections were investigated in the same way. The concentration of airborne sporangia in the vegetative cycle was also examined, as was the virulence of the different P. viticola populations detected. The epidemiological characteristics of the fungus differed depending on the degree of isolation of the vineyard, the airborne spore concentration, and on whether the attack was primary or secondary. Strong isolation was associated with the appearance of dominant fungal races and, therefore, reduced populational diversity.

• Journal of Microbiology and Biotechnology
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• v.33 no.3
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• pp.329-338
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• 2023

Enterohemorrhagic Escherichia coli (EHEC) is a foodborne pathogen that produces attaching and effacing lesions on the large intestine and causes hemorrhagic colitis. It is primarily transmitted through the consumption of contaminated meat or fresh produce. Similar to other bacterial pathogens, antibiotic resistance is of concern for EHEC. Furthermore, since the production of Shiga toxin by this pathogen is enhanced after antibiotic treatment, alternative agents that control EHEC are necessary. This study aimed to discover alternative treatments that target virulence factors and reduce EHEC toxicity. The locus of enterocyte effacement (LEE) is essential for EHEC attachment to host cells and virulence, and most of the LEE genes are positively regulated by the transcriptional regulator, Ler. GrlA protein, a transcriptional activator of ler, is thus a potential target for virulence inhibitors of EHEC. To identify the GrlA inhibitors, an in vivo high-throughput screening (HTS) system consisting of a GrlA-expressing plasmid and a reporter plasmid was constructed. Since the reporter luminescence gene was fused to the ler promoter, the bioluminescence would decrease if inhibitors affected the GrlA. By screening 8,201 compounds from the Korea Chemical Bank, we identified a novel GrlA inhibitor named Grlactin [3-[(2,4-dichlorophenoxy)methyl]-4-(3-methylbut-2-en-1-yl)-4,5-dihydro-1,2,4-oxadiazol-5-one], which suppresses the expression of LEE genes. Grlactin significantly diminished the adhesion of EHEC strain EDL933 to human epithelial cells without inhibiting bacterial growth. These findings suggest that the developed screening system was effective at identifying GrlA inhibitors, and Grlactin has potential for use as a novel anti-adhesion agent for EHEC while reducing the incidence of resistance.

• The Plant Pathology Journal
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• v.22 no.3
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• pp.215-221
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• 2006

Gibberella zeae (anamorph: Fusarium graminearum) is an important pathogen of cereal crops. This fungus produces a broad range of secondary metabolites, including polyketides such as aurofusarin (a red pigment) and zearalenone (an estrogenic mycotoxin), which are important mycological characteristics of this species. A screen of G. zeae insertional mutants, generated using a restriction enzyme-mediated integration (REMI) procedure, led to the isolation of a mutant (Z43R606) that produced neither aurofusarin nor zearalenone yet showed normal female fertility and virulence on host plants. Outcrossing analysis confirmed that both the albino and zearalenone-deficient mutations are linked to the insertional vector in Z43R606. Molecular characterization of Z43R606 revealed a deletion of at least 220 kb of the genome at the vector insertion site, including the gene clusters required for the biosynthesis of aurofusarin and zearalenone, respectively. A re-creation of the insertional event of Z43R606 in the wild-type strain demonstrated that the 220-kb deletion is responsible for the phenotypic changes in Z43R606 and that a large region of genomic DNA can be efficiently deleted in G. zeae by double homologous recombination. The results showed that 52 putative genes located in the deleted genomic region are not essential for phenotypes other than the production of both aurofusarin and zearalenone. This is the first report of the molecular characterization of a large genomic deletion in G. zeae mediated by the REMI procedure.