• IMMUNE NETWORK
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• v.14 no.2
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• pp.100-106
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• 2014

Infection with invasive Shigella species results in intestinal inflammation in humans but no symptoms in adult mice. To investigate why adult mice are resistant to invasive shigellae, 6~8-week-old mice were infected orally with S. flexneri 5a. Shigellae successfully colonized the small and large intestines. Mild cell death was seen but no inflammation. The infected bacteria were cleared 24 hours later. Microarray analysis of infected intestinal tissue showed that several genes that are involved with the sphingosine-1-phosphate (S1P) signaling pathway, a lipid mediator which mediates immune responses, were altered significantly. Shigella infection of a human intestinal cell line modulated host S1P-related genes to reduce S1P levels. In addition, co-administration of S1P with shigellae could induce inflammatory responses in the gut. Here we propose that Shigella species have evasion mechanisms that dampen host inflammatory responses by lowering host S1P levels in the gut of adult mice.

• The Korean Journal of Mycology
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• v.51 no.3
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• pp.179-190
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• 2023

Grapevine downy mildew, caused by Plasmopara viticola, significantly damages vineyards and is one of the most devastating diseases affecting cultivated grapes worldwide. In this study, we characterized the phenotypic and molecular traits of 11 P. viticola isolates from four grape-growing regions in South Korea. Additionally, we investigated the diversity of pathogenicity among these isolates and conducted an assay to evaluate the response of grape cultivars to P. viticola infection. Lemon-shaped sporangia were identified in the collected isolates, which released zoospores into the suspension at room temperature. Within a few hours of inoculation, the zoospores developed germ tubes. We tested 11 P. viticola isolates for pathogenicity in 845 grape cultivars to screen for grape host resistance to downy mildew infection. Among the tested isolates, JN-9 showed the highest virulence. Grape cultivars displayed varying phenotypic reactions to P. viticola infection: approximately 7% were highly susceptible, 41% were susceptible, 20% were moderately susceptible, 8% were resistant, and 24% exhibited extreme resistance. Phylogenetic analysis based on four genomic regions (internal transcribed spacer 1 [ITS1], actin, beta-tubulin, and cytochrome c oxidase II) revealed a close evolutionary relationship among all the Korean isolates, forming a single monophyletic lineage. Notably, these isolates showed greater similarity to European isolates than to American isolates. This comprehensive study contributes to a deeper understanding of the identity and behavior of P. viticola, which is crucial for developing effective resistance strategies against this pathogen in grape cultivars cultivated in South Korea.

• IMMUNE NETWORK
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• v.20 no.3
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• pp.25.1-25.13
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• 2020

Acinetobacter baumannii is known for its multidrug antibiotic resistance. New approaches to treating drug-resistant bacterial infections are urgently required. Cathelicidin-related antimicrobial peptide (CRAMP) is a murine antimicrobial peptide that exerts diverse immune functions, including both direct bacterial cell killing and immunomodulatory effects. In this study, we sought to identify the role of CRAMP in the host immune response to multidrug-resistant Acinetobacter baumannii. Wild-type (WT) and CRAMP knockout mice were infected intranasally with the bacteria. CRAMP^-/- mice exhibited increased bacterial colony-forming units (CFUs) in bronchoalveolar lavage (BAL) fluid after A. baumannii infection compared to WT mice. The loss of CRAMP expression resulted in a significant decrease in the recruitment of immune cells, primarily neutrophils. The levels of IL-6 and CXCL1 were lower, whereas the levels of IL-10 were significantly higher in the BAL fluid of CRAMP^-/- mice compared to WT mice 1 day after infection. In an in vitro assay using thioglycollate-induced peritoneal neutrophils, the ability of bacterial phagocytosis and killing was impaired in CRAMP^-/- neutrophils compared to the WT cells. CRAMP was also essential for the production of cytokines and chemokines in response to A. baumannii in neutrophils. In addition, the A. baumannii-induced inhibitor of κB-α degradation and phosphorylation of p38 MAPK were impaired in CRAMP^-/- neutrophils, whereas ERK and JNK phosphorylation was upregulated. Our results indicate that CRAMP plays an important role in the host defense against pulmonary infection with A. baumannii by promoting the antibacterial activity of neutrophils and regulating the innate immune responses.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.2100-2105
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• 2015

Probiotic bacteria must have not only tolerance against bile salt but also no genes for antibiotic resistance. Leuconostoc citreum is a dominant lactic acid bacterium in various fermented foods, but it is not regarded as a probiotic because it lacks bile salt resistance. Therefore, we aimed to construct a bile salt-resistant L. citreum strain by transforming it with a bile salt hydrolase gene (bsh). We obtained the 1,001 bp bsh gene from the chromosomal DNA of Lactobacillus plantarum and subcloned it into the pCB4170 vector under a constitutive P710 promoter. The resulting vector, pCB4170BSH was transformed into L. citreum CB2567 by electroporation, and bile salt-resistant transformants were selected. Upon incubation with glycodeoxycholic acid sodium salt (GDCA), the L. citreum transformants grew and formed colonies, successfully transcribed the bsh gene, and expressed the BSH enzyme. The recombinant strain grew in up to 0.3% (w/v) GDCA, conditions unsuitable for the host strain. In in vitro digestion conditions of 10 mM bile salt, the transformant was over 67.6% viable, whereas only 0.8% of the host strain survived.

• The Plant Pathology Journal
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• v.32 no.1
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• pp.47-52
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• 2016

Cotton leaf curl is devastating disease of cotton characterized by leaf curling, vein darkening and enations. The disease symptoms are induced by DNA satellite known as Cotton leaf curl Multan betasatellite (CLCuMuB), dominant betasatellite in cotton but another betasatellite known as Chili leaf curl betasatellite (ChLCB) is also found associated with the disease. Grafting experiment was performed to determine if host plant resistance is determinant of dominant population of betasatellite in cotton (several distinct strains of CLCuMuB are associated with the disease). Infected scion of Gossypium hirsutum collected from field (the source) was grafted on G. arboreum, a diploid cotton species, resistant to the disease. A healthy scion of G. hirsutum (sink) was grafted at the top of G. arboreum to determine the movement of virus/betasatellite to upper susceptible scion of G. hirsutum. Symptoms of disease appeared in the upper scion and presence of virus/betasatellite in the upper scion was confirmed via molecular techniques, showing that virus/betasatellite was able to move to upper scion through resistant G. arboreum. However, no symptoms appeared on G. arboreum. Betasatelites were cloned and sequenced from lower scion, upper scion and G. arboreum which show that the lower scion contained both CLCuMuB and ChLCB, however only ChLCB was found in G. arboreum. The upper scion contained CLCuMuB with a deletion of 78 nucleotides (nt) in the non-coding region between Arich sequence and ${\beta}C1$ gene and insertion of 27 nt in the middle of ${\beta}C1$ ORF. This study may help in investigating molecular basis of resistance in G. arboreum.

• The Plant Pathology Journal
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• v.37 no.4
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• pp.375-388
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• 2021

Rice yellow mottle virus (RYMV) is the most harmful virus that affects irrigated and lowland rice in Africa. The RBe24 isolate of the virus is the most pathogenic strain in Benin. A total of 79 genotypes including susceptible IR64 (Oryza sativa) and the resistant TOG5681 (O. glaberrima) as checks were screened for their reactions to RBe24 isolate of RYMV and the effects of silicon on the response of host plants to the virus investigated. The experiment was a three-factor factorial consisting of genotypes, inoculation level (inoculated vs. non-inoculated), and silicon dose (0, 5, and 10 g/plant) applied as CaSiO₃ with two replications and carried out twice in the screen house. Significant differences were observed among the rice genotypes. Fifteen highly resistant and eight resistant genotypes were identified, and these were mainly O. glaberrima. Silicon application did not affect disease incidence and severity at 21 and 42 days after inoculation (DAI); it, however, significantly increased plant height of inoculated (3.6% for 5 g CaSiO₃/plant and 6.3% for 10 g CaSiO₃/plant) and non-inoculated (1.9% for 5 g CaSiO₃/plant and 4.9% for 10 g CaSiO₃/plant) plants at 42 DAI, with a reduction in the number of tillers (12.3% for both 5 and 10 g CaSiO₃/plant) and leaves (26.8% for 5 g CaSiO₃/plant and 28% for 10 g CaSiO₃/plant) under both inoculation treatments. Our results confirm O. glaberrima germplasm as an important source of resistance to RYMV, and critical in developing a comprehensive strategy for the control of RYMV in West Africa.

• Genomics & Informatics
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• v.21 no.3
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• pp.42.1-42.23
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• 2023

Mycobacterium tuberculosis (Mtb) is the causative agent of tuberculosis, one of the most deadly infections in humans. The emergence of multidrug-resistant and extensively drug-resistant Mtb strains presents a global challenge. Mtb has shown resistance to many frontline antibiotics, including rifampicin, kanamycin, isoniazid, and capreomycin. The only licensed vaccine, Bacille Calmette-Guerin, does not efficiently protect against adult pulmonary tuberculosis. Therefore, it is urgently necessary to develop new vaccines to prevent infections caused by these strains. We used a subtractive proteomics approach on 23 virulent Mtb strains and identified a conserved membrane protein (MmpL4, NP_214964.1) as both a potential drug target and vaccine candidate. MmpL4 is a non-homologous essential protein in the host and is involved in the pathogen-specific pathway. Furthermore, MmpL4 shows no homology with anti-targets and has limited homology to human gut microflora, potentially reducing the likelihood of adverse effects and cross-reactivity if therapeutics specific to this protein are developed. Subsequently, we constructed a highly soluble, safe, antigenic, and stable multi-subunit vaccine from the MmpL4 protein using immunoinformatics. Molecular dynamics simulations revealed the stability of the vaccine-bound Tolllike receptor-4 complex on a nanosecond scale, and immune simulations indicated strong primary and secondary immune responses in the host. Therefore, our study identifies a new target that could expedite the design of effective therapeutics, and the designed vaccine should be validated. Future directions include an extensive molecular interaction analysis, in silico cloning, wet-lab experiments, and evaluation and comparison of the designed candidate as both a DNA vaccine and protein vaccine.

• Journal of the Korean Society of Tobacco Science
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• v.22 no.1
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• pp.7-12
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• 2000

This study was conducted to identify useful germ plasm sources of resistance to black shank, Phytophthora. parasitica, in Nicotiana tabacum and to characterize the interaction between host plant resistance to black shank and pathogen variability. The six internationally represen-tative set of black shank resistant and susceptible tobacco genotypes and four Korean lines were tested for black shank on the field infested with P. parasitica at Chonju Experiment Station, Korea Ginseng & Tobacco Research Institute during the 1992～1997 growing season. Almost 100% of the plants of the most susceptible genotype, Ky 14, did not survive. Complete survival of (MS Ky 14 x L8)F$_1$, which is resistant to reee 0 and ausceptible to reace 1, suggests that Race 0 is the only race of the P. parasitica in this bield. Coker 371 Gold and Beinhart 1000-1 having excellent resistance to black shank could be recommended as the parental cultivars for improving black shank resistance in Korea. Data of the diseased plants showed that KB 101 and KB 110 were medium high resistant and KB 103 and KB 108 being low resistant to black shank.

• Tuberculosis and Respiratory Diseases
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• v.44 no.3
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• pp.470-478
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• 1997

Background : Phagocytosis is probably the first step for mycobacteria to be virulent in host because virulent strains are more readily phagocytosed by macrophage than attenuated strains. According to the traditional concept, multi-drug resistant strains have been regarded as less virulent. However, this concept has been challenged, since recent studies(reported) showed that the degree of virulence and drug-resistance is not related. The purpose of this study is to evaluate whether the phagocytic activity of M.tuberculosis by peripheral blood mononuclear cells(PBMC) is different according to drug-resistance or host factor. To evaluate this, we estimated the difference of phagocytic activity of drug-resistant and drug-sensitive M.tuberculosis and also estimated the phagocytic activity of PBMC from intractable tuberculosis patients and healthy controls. Methods : PBMC from ten intractable tuberculosis patients and twelve healthy control, and three different strains of heat-killed M.tuberculosis, ie, ADS(all drug sensitive), MDR(multi-drug resistant), and ADR(all drug resistant) were used. After incubation of various strains of M.tuberculosis with PBMC, the phagocytic activity was evaluated by estimating proportion of PBMC which have phagocytosed M.tuberculosis. Results : Drug-resistant strains of M.tuberculosis were phagocytosed easily than drug sensitive strains(Percentage of PBMC phagocytosed M.tuberculosis in healthy control : ADS : $32.3{\pm}2.9%$, ADR : $49.6{\pm}3.4%$, p = 0.0022, Percentage of PBMC phagocytosed M.tuberculosis in intractable tuberculosis patients : ADS : $34.9{\pm}3.6%$, ADR : $50.7{\pm}4.5%$, p = 0.0069). However, there was no difference in phagocytic activity of PBMC from healthy control and intractable tuberculosis patients. Conclusion : Drug-resistant strains of M.tuberculosis were phagocytosed easily than drug sensitive strains and host factors does not seems to influence the phagocytosis of M.tuberculosis.

• Current Research on Agriculture and Life Sciences
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• v.32 no.3
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• pp.111-117
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• 2014

Phytophthora blight caused by Phytophthora capsici Leonian is a dangerous disease threatening pepper growers worldwide. The efficacy of chemical control is generally low as the pathogen is soil-borne and rapidly spread by zoospores during the rainy season. Thus, based on the demand for resistant varieties, various good resistant sources, such as CM334, AC2258, and PI201234, have been reported and their inheritance of resistance studied by many different authorities. However, the mode of inheritance remains unclear, as 1 or 2 independent dominant genes, 3 genes, or multiple genes have all been reported as responsible for resistance. Recently, QTL mappings of the gene factors for resistance have been reported, and molecular markers for resistance used in breeding programs. With the release of many resistant commercial hybrid cultivars, differentiation of pathotypes of the pathogen is attracting interest among breeders and plant pathologists. Various authorities have already classified the pathogen strains into different races according to the inter-action between resistant host plants, including the source of resistance, such as CM334 and PI201234, and resistant commercial varieties and P. capsici isolates. However, no standard differential host sets have yet been established, so the results are good only for the pathogen strains used in the experiments. Thus, for breeding varieties with durable resist-ance, it is important to introduce resistance from different sources and use diverse local pathogen strains collected in the target area for distribution in a breeding program.