• Journal of Life Science
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• v.32 no.2
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• pp.94-100
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• 2022

Chronic infection by hepatitis B virus (HBV) greatly increases the risk for liver cirrhosis and hepatocellular carcinoma (HCC). The outcome of HBV infection is shaped by the complex interplay of the mode of transmission, host genetic factors, viral genotype, adaptive mutations, and environmental factors. The pregenomic RNA transcription of HBV for their replication is regulated by the core promoter activation. Core promoter mutations have been the reason for acute liver failure and are associated with HCC development. We obtained HBV genes from a patient in Myanmar who was infected with HBV and identified gene variations in the core promoter region. For measuring the relative transactivation activity of the core promoter, we prepared the core-promoter reporter construct. Among the gene variations of the core promoter, the mutations of C1731T and G1806A were associated with increase in the transactivation of the HBV core promoter. Through computer analysis for searching for a tentative transcription factor binding site, we showed that the mutations of C1713T and G1806A newly created C/EBPβ and XBP1-responsive elements of the core promoter, respectively. The ectopic expression of C/EBPβ largely increased the HBV core promoter containing the C1713T mutation and that of XBP1 activated the M95 promoter containing the G1806A mutation. Our efforts to treat and prevent HBV infections are hampered by the emergence of drug-resistant mutations and vaccine-escape mutations. Our results provide the biological properties and clinical significance of specific HBV core promoter mutations.

• Research in Plant Disease
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• v.28 no.1
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• pp.19-25
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• 2022

Ginseng is an important medicinal plant cultivated in East Asia for thousands of years. It is typically cultivated in the same field for 4 to 6 years and is exposed to a variety of pathogens. Among them, ginseng root rot is the main reason that leads to the most severe losses. In this study, endophytic bacteria were isolated from healthy ginseng, and endophytes with antagonistic effect against ginseng root rot pathogens were screened out. Among the 17 strains, three carried antagonistic effect, and were resistant to radicicol that is a mycotoxin produced by ginseng root rot pathogens. Finally, Bacillus velezensis CH-15 was selected due to excellent antagonistic effect and radicicol resistance. When CH-15 was inoculated on ginseng root, it not only inhibited the mycelial growth of the pathogen, but also inhibited the progression of disease. CH-15 also carried biosynthetic genes for bacillomycin D, iturin A, bacilysin, and surfactin. In addition, CH-15 culture filtrate significantly inhibited the growth and conidial germination of pathogens. This study shows that endophytic bacterium CH-15 had antagonistic effect on ginseng root rot pathogens and inhibited the progression of ginseng root rot. We expected that this strain can be a microbial agent to suppress ginseng root rot.

• Journal of Life Science
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• v.33 no.1
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• pp.91-101
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• 2023

Bacteriocins are antimicrobial peptides synthesized on ribosomes, produced by bacteria, that inhibit the growth of similar or closely related bacterial strains. Since the discovery of nisin, many bacteriocins with unique structures and various modes of antibacterial activity have been described, and genes encoding production, secretion, and immunity have been reported. Nisin is one of the bacteriocins applied in cheese, liquid eggs, sauces and canned foods. Many of the bacteriocins of the genus Bacillus belong to lantibiotics, which are modified peptides after translation. Other genus Bacillus also produce many non-lantibiotic bacteriocins. Bacteriocins of the genus Bacillus are sometimes becoming more important because of their broader antibacterial spectrum. Bacteriocins are considered attractive compounds in the food and pharmaceutical industries to prevent food spoilage and growth of pathogenic bacteria. Bacteriocins can be used as biological preservatives in a variety of ways in the food system. Biopreservation refers to extending shelf life and improving safety of foods using microorganisms and/or their metabolites. The demand for new antimicrobial compounds has generated great interest in new technologies that can improve food microbiological safety. Applications of bacteriocins are expanding from food to human health. Today, many researchers are shifting their interest in bacteriocins from food preservation to the treatment of bacteria that cause infections and antibiotic-resistant diseases. This exciting new era in bacteriocin research will undoubtedly lead to new inventions and new applications. In this review, we summarize the various properties and applications of bacteriocins produced by the genus Bacillus.

• Korean Journal of Pharmacognosy
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• v.54 no.2
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• pp.72-79
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• 2023

MRSA is Staphylococcus aureus resistant to β-lactam antibiotics, and is a worldwide infectious disease. Even with the discovery of new antibiotics, resistance develops rapidly, so new alternatives are needed. Jakyakgamcho-tang (JGT) is a combination of Jakyak and Gamcho, and has been mainly used as an antispasmodic and analgesic in oriental medicine. This study was conducted to find out whether there is an effect on MRSA in relation to the anti-inflammatory effect of JGT and the antibacterial effect of Jakyak and Gamcho found in previous studies. In this study, in order to investigate the antibacterial activity of JGT and the combined effect of existing antibiotics, after extracting JGT with 70% EtoH, the disc diffusion method, minimum inhibitory concentration (MIC), drug combination effect (FICI), and time-kill analysis (Time-kill assay), metabolic inhibition, Western blot and qRT-PCR analysis were used to confirm the antibacterial activity mechanism of MRSA of JGT. As a result of the experiment, all of MRSA showed antibacterial activity in JGT's disc diffusion method, and the MIC was 250-1000 ㎍/mL. When existing antibiotics and JGT were combined with drugs, most had synergy or partial synergy. In addition, it was confirmed that the degree of bacterial growth was suppressed over time when simultaneous administration for 24 hours. JGT showed a synergistic effect when administered together with the ATPase-inhibitor DCCD, suggesting that it affected the inhibition of ATPase. As a result of observing the expression of PBP2a, and hla protein in the JGT-treated group and the untreated control group through wstern blot, it was confirmed that the protein expression of the JGT-treated group was significantly suppressed, and the expression levels of mecA, mecR1 and hla genes were also suppressed during JGT treatment. was observed by qRT-PCR. Combining the results of the experiment, it can be seen that JGT has antibacterial activity in MRSA, and when combined with existing antibiotics, the effect was increased compared to treatment with the drug alone. This suggests that JGT can be an alternative to treatment for antibiotic resistance of MRSA.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.269-269
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• 2022

Pre-harvest sprouting (PHS) on rice panicles is getting problematic in recent several years in Korea due to climate changes such as high temperature and more frequent typhoons during harvesting season. PHS negatively affects grain quality severely and also yield. Genetic improvement of Korean varieties (Oryza sativa ssp. japonica) through a marker assisted-backcross breeding (MAB) with the known PHS resistant genes must be one of ideal solutions. However, the final breeding products of MAB occasionally exhibit unwanted traits, especially the cross between genetically distant parents. This might be caused by linkage drag and/or presence of the gene-unlinked donor introgressions, resulting that the final products could not be released to the farmers. The major PHS resistance gene, Sdr4 (Seed dormancy 4) originated from an indica cultivar, Kasalath was selected as a donor gene. In order to avoid unexpected phenotypes in the breeding products, we performed a precision marker-based breeding (PMBB) consisting of foreground, recombinant, and background selections (FS, RS, and BS) which aim to develop 'single small introgression lines' (~100 kb introgression). Korean varieties (Ilpum and Gopum) were crossed with Kasalath. We developed Sdr4-allele specific markers for FS and a set of polymorphic flanking markers near the Sdr4 (-350kb and +420kb) for RS. To minimize linkage drag, the small introgression (< 125kb) containing Sdr4 was selected in Ilpum background (BC₂F₄) through 1^st RS with ~1,200 F₂ or BC₁F₂ plants (one side trimmed) and then 2^nd RS with ~1,000 progenies from the 1^st RS selected plants (another side trimmed). After RS, the selected lines were genotyped by using Infinium 7K SNP chip to detect other donor introgressions and the lines were backcrossed. Currently BS is on-going from the backcross-derived progenies with BS markers to remove residual introgressions. During the PMBB process, genetic effect of Sdr-4-Kasalath allele was confirmed in Ilpum and Gopum backgrounds by PHS phenotyping using the segregating BC₂F₃ or BC₁F₄ materials. The Sdr4 PMBB lines in Ilpum background (< 125kb introgression) will be valuable genetic resources to improve PHS resistance in modem popular temperate japonica varieties.

• Research in Plant Disease
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• v.30 no.2
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• pp.199-205
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• 2024

Erwinia amylovora, a causal pathogen of fire blight, has been continuously inducing damage to the apple and pear trees in South Korea since 2015. Farmers apply antibiotics during blooming season to prevent the fire blight. However, continuous use of antibiotics can induce the emergence of resistant bacteria, which consequently reduces control efficacy. In this study, we assessed the minimal inhibitory concentration (MIC) of streptomycin, using a total of 361 E. amylovora isolates that were collected from the six provinces of South Korea from 2019 to 2023. As a result, the MIC of streptomycin ranged from 0.5 to 4 ㎍/ml and the strA-strB genes were not identified from the isolates. The MIC was higher in the isolates from Gyeonggi-do, Gangwondo, and Chungcheongbuk-do compared to those from other three provinces. These results may bring broad attention to the use of streptomycin and aid in developing a management protocol for the occurrence of fire blight in South Korea.

• Research in Plant Disease
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• v.29 no.4
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• pp.363-376
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• 2023

Alternaria black spots or blights in cruciferous crops have been devastating diseases worldwide and led to economic losses in broccoli, Chinese cabbage, kale, radish, rapeseed, etc. These diseases are caused by different Alternaria spp., including A. brassicae, A. brassicicola and A. raphani transmitted from infected seeds or insect vectors. Efforts to excavate disease resistance traits of cruciferous crops against Alternaria black spots or blights have been demonstrated. Genetic resource of disease resistance was investigated in the wild relatives of cruciferous crops, and different cultivars were screened under different inoculation conditions. Development of the disease-resistant lines against Alternaria black spots or blights was also tried via genetic transformation of the cruciferous crops using diverse plant defence-associated genes. Plant immunity activated by pre-treatment with chemicals, i. e. β-amino-n-butyric acid and melatonin, was suggested for reducing Alternaria black spots or blights in cruciferous crops. The disease resistance traits have also been evaluated in model plant Arabidopsis originating from different habitats. Various plant immunity-related mutants showing different disease responses from wild-type Arabidopsis provided valuable information for managing Alternaria black spots or blights in cruciferous crops. In particular, redox regulation and antioxidant responses altered in the Alternaria-infected mutants were discussed in this review.

• Biomolecules & Therapeutics
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• v.32 no.4
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• pp.481-491
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• 2024

Paxlovid is the first approved oral treatment for coronavirus disease 2019 and includes nirmatrelvir, a protease inhibitor targeting the main protease (M^pro) of SARS-CoV-2, as one of the key components. While some specific mutations emerged in M^pro were revealed to significantly reduce viral susceptibility to nirmatrelvir in vitro, there is no report regarding resistance to nirmatrelvir in patients and animal models for SARS-CoV-2 infection yet. We recently developed xenograft tumors derived from Calu-3 cells in immunodeficient mice and demonstrated extended replication of SARS-CoV-2 in the tumors. In this study, we investigated the effect of nirmatrelvir administration on SARS-CoV-2 replication. Treatment with nirmatrelvir after virus infection significantly reduced the replication of the parental SARS-CoV-2 and SARS-CoV-2 Omicron at 5 days post-infection (dpi). However, the virus titers were completely recovered at the time points of 15 and 30 dpi. The virus genomes in the tumors at 30 dpi were analyzed to investigate whether nirmatrelvir-resistant mutant viruses had emerged during the extended replication of SARS-CoV-2. Various mutations in several genes including ORF1ab, ORF3a, ORF7a, ORF7b, ORF8, and N occurred in the SARS-CoV-2 genome; however, no mutations were induced in the M^pro sequence by a single round of nirmatrelvir treatment, and none were observed even after two rounds of treatment. The parental SARS-CoV-2 and its sublineage isolates showed similar IC₅₀ values of nirmatrelvir in Vero E6 cells. Therefore, it is probable that inducing viral resistance to nirmatrelvir in vivo is challenging differently from in vitro passage.

• Journal of Korean Society of Forest Science
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• v.39 no.1
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• pp.47-56
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• 1978

This study aimed to know difference in freezing resistance among different clonal seedlings or different seed source seedlings of Cryptomeria japonica which has been selected where extreme cold prevails in Korea and Japan. The freezing resistance of three 12-50 year old trees was also measured in the experiment. The freezing resistance was measured in different tissue parts: mainly leaf, cambiam and xylem, at three different collection dates in two different collection places during the winter of 1977-1978. The following results and discussions were made: 1. The clonal difference in freezing resistance of Cryptomeria japonica was $9^{\circ}$ to $15^{\circ}C$ in maximum according to the collection place. However, the clonal difference in freezing resistance was not related to the difference in climatic conditions where the parent tree have been growing. This impiled that the natural selection of cold resistant genes in Cryptomeria japonica has not reached its evolutional equilibrium yet since most of the Cryptomeria forest has been established by artificial regeneration. 2. The difference in freezing resistance among leaf, cambium and xylem was not apparent except that leaf of several clones showed higher freezing resistance than cambium or xylem when they collected at mid-winter. The least freezing resistant tissue part, thought its freezing resistance was not measure in all clones and all temperatures were appeared in the apical buds. The new shoot growth was observed in the next spring with being replaced by its dormant or adventitious bud growth when the apical bud was injured dy cold during winter. 3. The freezing resistance of leaf, cambium and xylem was shown high enough so that freezing resistance Cryptomeria clones in this experiment were supposed to be able to survive in cold winter conditions at the middle part of Korea. However, it was reported that the most susceptible tissue part to winter injury was the basal stem, but of which freezing resistance was not-measured in this experiment. Several silvicultural methods for prevention of Cryptomeria seedlings from cold damage were discussed in literature.

• Tuberculosis and Respiratory Diseases
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• v.54 no.4
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• pp.367-377
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• 2003

Background : It is well known that only 10% of those infected with Mycobacterium tuberculosis actually develop clinical disease, indicating the existence of host genetic factors regulating disease expression. In this study, we investigated HLA-DRB1 and -DQB1 gene polymorphisms in Korean patients with pulmonary tuberculosis (PTB). Methods : HLA-DRB1 and -DQB1 gene polymorphisms were investigated in 67 PTB patients without previous treatment history, 38 drug-sensitive (DS) and 29 multidrug-resistant (MDR) cases, and 200 healthy controls. HLA-DRB1 typing was done using reverse SSO (sequence specific oligonucleotide) and PCR-SSCP (single strand conformational polymorphism) methods and DQB1 typing was done using PCR-RFLP (restriction fragment length polymorphism), PCR-SSCP and PCR-SSP (sequence specific primer) methods. Results : Among the PTB patients, MDR-TB cases showed frequencies of DRB1*0701 and *08032 increased by about two-fold compared to those of normal controls, and likewise for their associated DQB1 alleles, DQB1*0202 and *0601 (15.5% vs. 34.5%, p=0.01). The frequency of HLA-DQB1*0609 was significantly increased in PTB patients (4.0% vs. 14.9%, p=0.004), showing similar increases in both DS and MDR cases. There was also an association of HLA alleles with the clinical severity of the disease according to the extent of lung lesion. Significantly increased frequencies of DRB1*08032 (4.2% vs. 32.6%, p=0.007) and DQB1*0601 (12.5% vs. 34.9%, p=0.047) were observed in more advanced (moderately & far advanced/DS and far advanced/MDR), compared with less advanced (minimal/DS and moderately advanced/MDR) lung lesions. Although DRB1*0701, DQB1*0202 and DQB1*0609 showed significant increases in different subsets of the disease, these HLA alleles did not show consistent association with disease severity. Conclusion : HLA-DRB1*08032 and DQB1*0601 alleles were associated with genetic susceptibility to MDR-TB in Korean patients, and also with disease severity and progression of PTB.