• Korean Journal of Microbiology
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• v.48 no.4
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• pp.284-292
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• 2012

To date, chemical control remains the most common way to reduce beet armyworm (Spodoptera exigua) populations. However, this insect has become more tolerant or resistant to many chemical insecticides and the insect larvae usually hide inside hollow, tube-like leaves of host plant so they were difficult to kill by spraying insecticides. The use of viral and bacterial insecticide to solve these problems has not been successful because of their novel feeding habit. To overcome these problems, in this study, the biological characteristics and virulence of an entomopathogenic fungus isolated from the cadaver of larvae beet armyworm were investigated. Isolated entomopathogenic fungus was identified as Nomeraea rileyi (Farlow) Samson by morphological examinations and genetic identification using sequences of the ITS, ${\beta}$-tubulin gene and EF1-${\alpha}$ regions. This fungus was named as N. rileyi SDSe. Virulence tests against 3rd larvae of beet armyworm were conducted with various conidial suspensions from $1{\times}10^4$ to $10^8$ conidia/ml of N. rileyi SDSe in laboratory conditions. Mortality rate of beet armyworm showed from 20 to 54% and the virulence increased with increasing conidial concentrations. Although N. rileyi SDSe showed low mortality rate against beet armyworm, it is expected that N. rileyi SDSe will be used effectively in the integrated pest management programs against the beet armyworm.

• Journal of the korean academy of Pediatric Dentistry
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• v.33 no.3
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• pp.401-410
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• 2006

Dental caries results from localized demineralization of tooth enamel by acids of bacterial origin produced from the fermentation of dietary sugars. A group of related oral bacteria, collectively known as mutans streptococci, are implicated as the primary etiological agents of human caries. Within this group, Streptococcus mutans has been known as a causative agent for dental caries. As well as acid production yielding the demineralization of tooth enamel, adherence and colonization of S. mutans to the teeth are also important for their virulence Cell-surface fibrillar proteins, which mediate adherence to the salivary pellicle are virulence components of mutans streptococci, and primary candidates for a human caries vaccine. Here we report that the AgI/II gene from S. mutans GS-5 were cloned by PCR amplification of the bacterial chromosomal DNA and the integrity of cloned genes were confirmed by nucleotide sequencing. Sequence analyses showed the sequence alignment of 280 nucleotides between the cloned AgI/II and the reported sequence of S. mutans GS-5 showed the perfect match The cloned genes which signal nucleotide was truncated, were transferred into bacterial expression vector and the recombinant proteins were purified as His-tag fusion proteins In order to generate polyclonal antibodies against the recombinant proteins, AgI/II mr, some $100{\mu}g$ of the proteins was injected into mice three times. It can be used for an effective vaccine production to prevent dental caries caused by pathogenic S. mutans.

• Journal of Life Science
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• v.25 no.2
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• pp.136-150
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• 2015

Pseudomonas syringae pathovar tabaci is a plant pathogenic bacterium that causes wildfire disease in tobacco plants. In P. syringae pv. tabaci, PsyI, a LuxI-type protein, acts as an AHL synthase, while primary and secondary sequence analysis of PsyR has revealed that it is a homolog of the LuxR-type transcriptional regulator that responds to AHL molecules. In this study, using phenotypic and genetic analyses in P. syringae pv. tabaci, we show the effect of PsyR protein as a quorum-sensing (QS) transcriptional regulator. Regulatory effects of PsyR on swarming motility and production of siderophores, tabtoxin, and N-acyl homoserine lactones were examined via phenotypic assays, and confirmed by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Further qRT-PCR showed that PsyR regulates expression of these virulence genes in response to environmental signals. However, an upstream region of the gene was not bound with purified MBP-PsyR protein; rather, PsyR was only able to shift the upstream region of psyI. These results suggested that PsyR may be indirectly controlled via intermediate-regulatory systems and that auto-regulation by PsyR does not occur.

• Food Science of Animal Resources
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• v.36 no.6
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• pp.779-786
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• 2016

This study aimed to evaluate the prevalence of Listeria monocytogenes on livestock farms in Korea and determine their serotypes and genetic correlations. Twenty-five livestock farms in Korea (central: 15, south west: 7, south east: 3) were visited 2-3 times, and 2,018 samples (feces: 677, soil: 680, silage: 647, sludge: 14) were collected. Samples were enriched in LEB (Listeria enrichment broth) and Fraser broth media, and then plated on Palcam agar. The isolates were identified by PCR and 16S rRNA gene sequencing. Then, the sero-types, presence of virulence genes (actA, inlA, inlB, plcB, and hlyA), and antibiotic resistance were determined. Genetic correlations among the isolates were evaluated by analyzing the restriction digest pattern with AscI. Of the 2,018 samples, only 3 (0.15%) soil samples (FI-1-FI-3) from 1 farm in the south east region were positive for L. monocytogenes. Based on biochemical tests and multiplex PCR, the serotype of the isolates were 4ab (FI-1 and FI-3) and 3a (FI-2), which are not common in foodborne L. monocytogenes. The 3a sero-type isolate was positive for all tested virulence genes, whereas the 4ab serotype isolates were only positive for hlyA, actA, and inlA. The isolates were resistant to all 12 tested antibiotics, especially FI-3. The genetic correlations among the isolates were 100% for those of the same serotype and 26.3% for those of different serotypes. These results indicate that the prevalence of L. monocytogenes on livestock farms in Korea is low; however, the isolates are pathogenic and antibiotic resistant.

• Journal of Life Science
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• v.31 no.1
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• pp.28-36
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• 2021

Edwardsiella piscicida is a significant cause of hemorrhagic septicemia in fish and gastrointestinal infections in humans. Survival bacteria require specialized mechanisms to adapt to environmental fluctuations. Hence, to understand the mechanism through which E. piscicida senses and responds to environmental osmolarity changes, we determined the protein expression profile and physiological properties under various salinity conditions in this study. The OmpR protein is a part of the Env-ZOmpR two-component system that has been implicated in sensing salt stress in bacteria. However, the physiological role played by this protein in E. piscicida remains to be elucidated. Therefore, in this work, the function of the OmpR protein in response to salt stress was investigated. Phenotypic analysis revealed that, in the mutant, three of the biochemical phenotypes were different from the wild type, including, citrate utilization, hydrogen sulfide, and indole production. Introduction of the plasmid containing the entire ompR gene to the mutant strain returned it to its parental phenotype. The retarded growth rate also partially recovered. Furthermore, in our studies, OmpR was not found to be related to cell motility. Taken together, our results from the mutational analysis, the growth assay, MALDI-TOF MS, qRT-PCR, and the phenotype studies suggest that the OmpR of E. piscicida is implicated in osmoregulation, growth, expression of porins (ETAE_1826), virulence-related genes (EseC, EseD and EvpC), and certain genes of unknown function (ETAE_1540 and ETAE_2706).

• Molecules and Cells
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• v.39 no.5
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• pp.426-438
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• 2016

Plant disease resistance occurs as a hypersensitive response (HR) at the site of attempted pathogen invasion. This specific event is initiated in response to recognition of pathogen-associated molecular pattern (PAMP) and subsequent PAMP-triggered immunity (PTI) and effector-triggered immunity (ETI). Both PTI and ETI mechanisms are tightly connected with reactive oxygen species (ROS) production and disease resistance that involves distinct biphasic ROS production as one of its pivotal plant immune responses. This unique oxidative burst is strongly dependent on the resistant cultivars because a monophasic ROS burst is a hallmark of the susceptible cultivars. However, the cause of the differential ROS burst remains unknown. In the study here, we revealed the plausible underlying mechanism of the differential ROS burst through functional understanding of the Magnaporthe oryzae (M. oryzae) AVR effector, AVR-Pii. We performed yeast two-hybrid (Y2H) screening using AVR-Pii as bait and isolated rice NADP-malic enzyme2 (Os-NADP-ME2) as the rice target protein. To our surprise, deletion of the rice Os-NADP-ME2 gene in a resistant rice cultivar disrupted innate immunity against the rice blast fungus. Malic enzyme activity and inhibition studies demonstrated that AVR-Pii proteins specifically inhibit in vitro NADP-ME activity. Overall, we demonstrate that rice blast fungus, M. oryzae attenuates the host ROS burst via AVR-Pii-mediated inhibition of Os-NADP-ME2, which is indispensable in ROS metabolism for the innate immunity of rice. This characterization of the regulation of the host oxidative burst will help to elucidate how the products of AVR genes function associated with virulence of the pathogen.

• Korean Journal of Microbiology
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• v.49 no.4
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• pp.383-390
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• 2013

Lateral gene transfer (LGT) of genes from other bacteria into Vibrio cholerae is expectable because of the pronounced natural competence of the bacterium. In this study, quantitative aspects of LGT among the three species of Vibrio pathogenic to humans were characterized. Genome sequences of V. cholerae N16961, V. parahaemolyticus RIMD2210633, V. vulnificus CMCP6, and Escherichia coli K12 substrain MG1655 were analyzed to determine orthologous quartets of protein coding genes present in all four genomes. Phylogenetic analyses on the quartets were conducted to resolve vertical versus lateral patterns of gene polymorphisms based on congruence versus incongruence of phylogenetic trees. About 70% of the quartets could be resolved as either cohesive topology (75%) or LGT tree topologies (25%). The amount of LGT genes in Vibrio spp. appeared to be abnormally high for a genus and comparable to those of families. Patched distributions of LGT from different donors were observed on a chromosome. In the small chromosome of V. cholerae, physical linkages among LGT loci spanned half the length of the chromosome. Either accumulative selection for the donor alleles in LGT or presence of large-scale LGT events was hypothesized. These findings warrant further studies on the nature of donor-specificity of LGT alleles and its influence on evolution of Vibrio virulence to humans.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.20
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• pp.9027-9031
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• 2014

Background: Helicobacter pylori (H. pylori) infection is an established cause of peptic ulcers and gastric cancer. The aim of this study was to identify H. pylori genotypes and to examine their associations with geographical regions and gastritis, peptic ulcers and gastric cancer in Laos. Materials and Methods: A total of 329 Lao dyspeptic patients who underwent gastroscopy at Mahosot Hospital, Vientiane, Laos during December 2010 - March 2012 were enrolled. Two biopsy specimens (one each from the antrum and corpus) were obtained for CLO testing and only CLO test-positive gastric tissue were used to extract DNA. PCR and sequencing were identified for variants of the cagA and vacA genotypes. Results: Some 119 Laos patients (36.2%) were found to be infected with H. pylori including 83 with gastritis, 13 with gastric ulcers (GU), 20 with duodenal ulcers (DU) and 3 with gastric cancer. cagA was detected in 99.2%. East-Asian-type cagA (62%) and vacA s1c (64.7%) were predominant genotypes in Laos. vacA s1c-m1b was significantly higher in GU than gastritis (53.8% vs. 24.1%; P-value=0.04) whereas vacA s1a-m2 was significantly higher in DU than gastritis (40.0% vs. 16.9%; P-value=0.03). East-Asian-type cagA and vacA s1c were significantly higher in highland than lowland Lao (100% vs. 55.8%; P-value=0.001 and 88.2% vs. 61.5%, P-value=0.03 respectively). Conclusions: H. pylori is a common infection in Laos, as in other countries in Southeast Asia. The cagA gene was demonstrated in nearly all Laos patients, cagA and vacA genotypes being possible important factors in explaining H. pylori infection and disease outcomes in Laos.

• Journal of Food Hygiene and Safety
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• v.6 no.3
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• pp.147-155
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• 1991

Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin(ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxingeic E. coli from non pathogenic E. coli. The isolate of enterotoxigenlc E. coli was isolated from swine during 1989 year(from 5 to 10 month) in the Kyong-gi and Chung-Cheong provinces, and three strains(KM-4, KM-7 and KM-12) was selected from 189 isolates of ST producing E. coli. The detection of a ST produced of the isolated E. coli was performed by the infant mouse assay(IMA). This study was designed to know optimal conditions for the production of the ST and the molecular properties of plasmids of the enterotoxigenic E. coli. Amount of ST produced were the most at initial pH 8.5~9.0 of succinate salts medium culture. The cultural time of the same medium was accumulated the highest level of ST was at the 14 to 16 hours, and then stationary phase was at the 20 hours. From this experiment the KM-7 strain was selected among ST producing strains by IMA. Partial plasmid-curing experiment was done to select plasmid encoding for ST among other plasmids and then comparing the plasmid pattern of ST producing strain(KM-7) with those of other ST non-producing strains, it is found that ST gene exists on the about 80 Kbp plasmid. Each fragment of this plasmid digested with EcoRl was ligated to vector pBR 322 and transformed into E. coli K-12. A clone producing ST(eKT 53) was selected by IMA. The EcoRl digestion pattern of the isolated plasmid(pKD 37) from the ST producing clone it is indicated that the size of the inserted fragment in eKT 53 strain is 16 Kbp. The cultured supernatant of eKT 53 strain was positive result of ST production in IMA.

• Research in Plant Disease
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• v.21 no.2
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• pp.99-102
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• 2015

Cucumber mosaic virus (CMV) is one of the most destructive viruses in chilli pepper. An isolate of CMV was obtained from the chilli pepper cv. Chungyang showing top necrosis symptom in 2013 and designated as CMV-GTN. CMV-GTN was compared with the well-characterized isolate, CMV-Ca-P1, by investigating their amino acid sequences of the coat protein (CP) and biological reactions in several host plants. The CP of CMV-Ca-P1 composed of 217 amino acids but that of CMV-GTN composed of 218 amino acids by including additional valine in the $57^{th}$ amino acid position. Amino acid sequence similarity of the CP gene among CMV-GTN and other CMV isolates recorded in the GeneBank database ranged from 96% to 99%. CMV-GTN was selected as a representative isolate to screen the resistance pepper cultivars to CMV because it was highly pathogenic to tomatoes and peppers upon biological assays. The virulence of CMV-GTN was tested on 135 pepper cultivars which has been bred in Korea and compared with that of CMV-Ca-P1. Only the cv. Premium was resistant and three cvs. Hot star, Kaiser, and Good choice were moderately resistant to CMV-GTN, whereas two cvs. Baerotta and Kaiser were resistant to CMV-Ca-P1.