• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.121-127
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• 2004

Amphotericin B (AmB), an amphipathic polyene macrolide, is an antifungal drug produced by Streptomyces nodosus. Recently, AmB has been shown to exert antiviral activity against rubella virus and human immunodeficiency virus by different mechanisms. In this study, we evaluated the antiviral effect of AmB against Japanese encephalitis virus (JEV) and investigated which step of the viral life cycle was inhibited by AmB to understand the mechanism of antiviral action of AmB. AmB reduced both plaque size and number in the infected cells in a dose-dependent manner. In addition, a 200-fold reduction of infectious virus titer was observed by treatment of infected cells with $5\mug/ml$ of AmB. AmB acted at the post virus-infection step, but not during adsorption of virus to host cells. Western blot analysis revealed that the accumulated level of JEV envelope protein dramatically decreased in the infected cells by treatment with $5-10\mug/ml$ of AmB. Our results indicate that AmB inhibits the replication of JEV at the postinfection step by interfering with viral replication and/or by inhibiting the synthesis of viral proteins.

• The Plant Pathology Journal
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• v.35 no.5
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• pp.417-424
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• 2019

Melanose, caused by Diaporthe citri, is one of severe diseases in citrus, a major economic resource in Jeju island. To reduce the usage amount of organic synthetic fungicide, bio-sulfur was tested as an alternative chemical to control citrus melanose in the present study. Direct antifungal activity of bio-sulfur against D. citri was determined through in vitro experiment using artificial nutrient media. Disease severity of melanose on bio-sulfur pretreated citrus leaves was lower than that on untreated ones. To illustrate the mechanism of disease suppression by bio-sulfur, infection structures were observed with a fluorescent microscope and a scanning electron microscope. In fluorescent microscopic observation, most conidia rarely germinated. In addition, hyphal growth on leaves pretreated with bio-sulfur was inhibited compared to that on untreated ones. In scanning electron microscope images of bio-sulfur pretreated leaves, surfaces of most conidia were shrunk while hyphae were morphologically changed and frequently branched. Such microscopic observations were also found for leaves pretreated with a commercial fungicide Dithianon. These results suggest that bio-sulfur may be used to control citrus melanose as an environment friendly alternative to organic synthetic fungicides

• Journal of Applied Biological Chemistry
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• v.52 no.3
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• pp.116-120
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• 2009

We found out the new method of the consortium for the environmental friendly agriculture by 8 kinds of the selected antagonistic rhizobacteria. This research involved composition of mutual complementary consortium by each antagonistic function such as production of antibiotic, siderophore, antifungal cellulase and insoluble phosphate solubilization. The consortium No.11 among composed consortium candidates showed the most pepper growth promoting activity and Phytophthora blight suppression on the in vivo pot test of red-pepper plant. The consortium No. 11 is combination of PGPR Bacillus subtilis AH18 and Bacillus licheniformis K11. B. subtilis AH18 and B. licheniformis K11 both could produce the auxin, antifungal ${\beta}$-glucannase and siderophore. Also, they had mechanism for solubilization of insoluble phosphate. But, B. licheniformis K11 could produce the antibiotic of iturin which was able to inhibit Phytophthora capsici. We confirmed complementary noncompetitive mutualism between B. subtilis AH18 and B. licheniformis K11 of the consortium No.11. The results came out through treatment of two strains co-culture, treatment of individual culture and co-treatment of two individual cultures for the growth and Phytophthora blight suppression of red-pepper. The treatment of two strains co-culture didn't show a synergic effect in comparing sole treatment on the pepper growth promotion and Phytophthora blight suppression. But, when the pots were treated simultaneously with co-treatment of two individual cultures, an synergic effect was seen in the growth promotion of roots, stem, leaves and suppressed Phytophthora blight on red-pepper in vivo pot test.

• Microbiology and Biotechnology Letters
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• v.36 no.4
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• pp.268-275
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• 2008

In order to develop the multi-functional rhizobacteria that can exert positive effect on the growth of plants growing in the coastal sand dune located along East Coast of Korea, rhizospheral bacteria of 11 different plants from this area were isolated 1,330 rhizobacteria. Among these, 23 strains were able to produce auxin and had spectrum of antagonism toward various phytopathogenic microbes. To know the mechanism of this antifungal activity, these 23 strains were subjected to further analyses; 19 strains of these produced siderophore as determined by color reaction on CAS-blue plate, 4 strains produced antifungal cellulase as judged by color change on CMC-Congo red plate, 17 strains were able to utilized insoluble phosphate salts, also determined by clear zone formation on PVK medium. Identification of the strain was assigned to all 23 strains by l6s rDNA sequence analysed, and all were identified to be in the genus of Bacillus and Pseudomonas. One strain of these, denoted Pseudomonas fluorescens IB4-14, showed ACC deaminase activity which is known to be involved in the resistance of environmental stress such as salt and drought. Also, P. fluorescens IB4-l4 showed the germination stimulation and roots growth promoting activity on the in vivo assay of Lysimachia mauritiana Lam. (spoonleaf yellow loosestrife).

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.301-310
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• 2010

Bacillus subtilis strains produce a broad spectrum of bioactive peptides. The lipopeptide surfactin belongs to one well-known class, which includes amphiphilic membrane-active biosurfactants and peptide antibiotics. Both the srfA promoter and the ComP-ComA signal transduction system are an important part of the factor that results in the production of surfactin. Bs-M49, obtained by means of low-energy ion implantation in wild-type Bs-916, produced significantly lower levels of surfactin, and had no obvious effects against R. solani. Occasionally, we found strain Bs-M49 decreased spore formation and the development of competence. Blast comparison of the sequences from Bs-916 and M49 indicate that there is no difference in the srfA operon promoter PsrfA, but there are differences in the coding sequence of the comA gene. These differences result in three missense mutations within the M49 ComA protein. RT-PCR analyses results showed that the expression levels of selected genes involved in competence and sporulation in both the wild-type Bs-916 and mutant M49 strains were significantly different. When we integrated the comA ORF into the chromosome of M49 at the amyE locus, M49 restored hemolytic activity and antifungal activity. Then, HPLC analyses results also showed the comA-complemented strain had a similar ability to produce surf actin with wild-type strain Bs-916. These data suggested that the mutation of three key amino acids in ComA greatly affected the biological activity of Bacillus subtilis. ComA protein 3D structure prediction and motif search prediction indicated that ComA has two obvious motifs common to response regulator proteins, which are the N-terminal response regulator receiver motif and the C-terminal helix-turn-helix motif. The three residues in the ComA N-terminal portion may be involved in phosphorylation activation mechanism. These structural prediction results implicate that three mutated residues in the ComA protein may play an important role in the formation of a salt-bridge to the phosphoryl group keeping active conformation to subsequent regulation of the expression of downstream genes.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.88-95
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• 2013

Candida biofilms are self-organized microbial communities growing on the surfaces of host tissues and medical devices. These biofilms have been displaying increasing resistance against conventional antifungal agents. The roots of Scutellaria baicalensis have been widely used for medicinal purpose throughout East Asia. The aim of the present study was to evaluate the effect of S. baicalensis aqueous extract upon the preformed biofilms of 10 clinical C. albicans isolates, and assess the mechanism of the antibiofilm activity. Its effect on preformed biofilm was judged using an XTT reduction assay and the metabolic activity of all tested strains were reduced ($57.7{\pm}17.3$%) at MIC values. The S. baicalenis extract inhibited (1, 3)-${\beta}$-D-glucan synthase activity. The effect of S. baicalensis on the morphology of C. albicans was related to the changes in growth caused by inhibiting glucan synthesis; most cells were round and swollen, and cell walls were densely stained or ruptured. The anticandidal activity was fungicidal, and the extract also arrested C. albicans cells at $G_0/G_1$. The data suggest that S. baicalensis has multiple fatal effects on target fungi, which ultimately result in cell wall disruption and killing by inhibiting (1, 3)-${\beta}$-D-glucan synthesis. Therefore, S. baicalensis holds great promise for use in treating and eliminating biofilm-associated Candida infections.

• Korean Journal of Soil Science and Fertilizer
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• v.41 no.1
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• pp.18-25
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• 2008

Pseudomonas sp. PRGB06, a bacterial strain isolated from diamondback moth (Plutella xylostella) gut, was examined for its plant growth promotion and biofertilizing traits. The bacteria growth was observed under various conditions of carbon sources, temperature, pH and salt concentrations. In addition, the mechanisms of antagonism and phosphate solubilization were investigated. The bacterial strain PRGB06, grew well using most of the tested carbon sources. The best growth was observed at $30^{\circ}C$ and pH 7. The inhibition of the pathogenic fungi was likely due to the volatile antifungal metabolite and ammonia gas produced by the bacteria. A significant positive relationship was found between the phosphate solubilization and acid production. When inoculated with PRGB06 in vitro and in gnotobiotic condition, red pepper and maize showed increase in root length, seedling vigor and dry bio-mass.

• Microbiology and Biotechnology Letters
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• v.33 no.3
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• pp.194-199
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• 2005

In oder to select an antifungal substance-producing antagonistic bacterium against Fusarium oxysporum casuing fusarium wilt on tomato, strain BK4 was isolated from local soil of Gyeoungbuk and was identified as Bacillus thuringiensis by 16s rDNA analysis, biochemical test, and Mcirolog TM 3.0 System. The antibiotic of B. thuringiensis BK 4 was highly produced at $30^{\circ}C$ in nutrient broth (pH 9.0). The crude antibiotic was even stable at $121^{\circ}C$ and more stable at slight alkalic condition than acid condition. It was also remained $50{\%}$ activity at pH 3.0. B. thuringiensis BK4 showed the inhibition of spore germination and the biocontrol ability against F. oxysporum causing fusarium wilt of tomato in vivo test. According to these results, B. thuringiensis BK4 was enough to use with a microbial agent for biocontrol against fusarium wilt.

• BMB Reports
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• v.34 no.1
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• pp.15-20
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• 2001

One of the innate immune reactions in invertebrates is the pro-phenoloxidase (pro-PO) activation system that is involved in the generation of superoxide, melanin synthesis, and the subsequent sequestration of foreign matter entering the hemocoel of the invertebrates. However, the molecular mechanism of this biological reaction is still obscure. To expand our understanding of the biological roles of the pro-PO activation system in invertebrates, we performed a yeast two-hybrid screening by using three regions of pro-PO as bait and a yeast two-hybrid cDNA library from Tenebrio molitor larvae as prey We isolated a novel partial cDNA clone that encodes a glycine-rich protein that interacted with the active phenoloxidase (termed phenoloxidase interacting protein, POIP). POIP consists of two domains: One is an N-terminal unique domain and the other is a C-terminal glycine-rich domain. The C-terminal glycine-rich domain showed sequential homology with those of insect antifungal proteins. Also, the yeast two-hybrid screen in a reverse orientation (using POIP as bait) yielded PO, suggesting that the PO-POIP interaction is specific. By using a 315 bP PCR fragment of the N-terminal unique region of POIP, we cloned the full-length cDNA of POIP from the Tenebruo cDNA library constructed by using E. coli injected larvae. The interaction analysis between PO, and a truncated fragment lacking the N-terminal unique region of POIP, indicated that the N-terminal unique region is necessary for interaction between PO and POIP. The expression level of the POIP mRNA is increased by bacterial injection into T. molitor larvae. This suggests that POIP might be engaged in the humoral defense reaction.

• Journal of Microbiology and Biotechnology
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• v.28 no.3
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• pp.482-490
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• 2018

Candida albicans infections are often problematic to treat owing to antifungal resistance, as such infections are mostly associated with biofilms. The ability of C. albicans to switch from a budding yeast to filamentous hyphae and to adhere to host cells or various surfaces supports biofilm formation. Previously, the ethanol extract from Paeonia lactiflora was reported to inhibit cell wall synthesis and cause depolarization and permeabilization of the cell membrane in C. albicans. In this study, the P. lactiflora extract was found to significantly reduce the initial stage of C. albicans biofilms from 12 clinical isolates by 38.4%. Thus, to assess the action mechanism, the effect of the P. lactiflora extract on the adhesion of C. albicans cells to polystyrene and germ tube formation was investigated using a microscopic analysis. The density of the adherent cells was diminished following incubation with the P. lactiflora extract in an acidic medium. Additionally, the P. lactiflora-treated C. albicans cells were mostly composed of less virulent pseudohyphae, and ruptured debris was found in the serum-containing medium. A quantitative real-time PCR analysis indicated that P. lactiflora downregulated the expression of C. albicans hypha-specific genes: ALS3 by 65% (p = 0.004), ECE1 by 34.9% (p = 0.001), HWP1 by 29.2% (p = 0.002), and SAP1 by 37.5% (p = 0.001), matching the microscopic analysis of the P. lactiflora action on biofilm formation. Therefore, the current findings demonstrate that the P. lactiflora ethanol extract is effective in inhibiting C. albicans biofilms in vitro, suggesting its therapeutic potential for the treatment of biofilm-associated infections.