• Research in Plant Disease
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• v.19 no.3
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• pp.208-215
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• 2013

3-(4-Hydroxyphenyl)-propionic acid (HPP) is a bacterial metabolite synthesized and released by an entomopathogenic bacterium Xenorhabdus nematophila K1. In this study, the control efficacy of HPP was tested against Phytophthora blight and anthracnose of red pepper plants. HPP suppressed mycelial growth of Phytophthora blight and anthracnose pathogens. Under natural sunlight condition, HPP maintained the antifungal activity on the diseases for at least twenty five days. The antifungal activity was not decreased even in the condition of soil-water. It was proved that HPP was able to penetrate the roots and travel upward of the red pepper plants. When HPP suspension was applied to soil rhizosphere before transplanting the red pepper seedlings or was regularly sprayed to the foliage of the plants with ten days interval, it resulted in significant reduction of the disease occurrences (Phytophthora blight and anthracnose) without any phytotoxicity. These results suggested that HPP can be developed to a systemic agrochemical against Phytophthora blight and anthracnose of red pepper plants.

• Microbiology and Biotechnology Letters
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• v.50 no.4
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• pp.488-500
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• 2022

Most fungal infections by opportunistic yeast pathogens such as Candida spp. are the major causes of morbidity and mortality in patients with lowered immune. Previous studies have reported that some strains of Candida secret secondary metabolites play an important role in the decreasing of immunity in the infected patient. In this study, 110 Candida spp. were isolated from different clinical specimens from Baghdad hospitals. Candida isolates were identified by conventional methods, they were processed for Candida speciation on CHROMagar. The results of identification were confirmed by internal transcribed spacer (ITS) sequencing. Phylogenetic trees were analyzed with reference strains deposited in GenBank. Antifungal susceptibility testing was evaluated by the disc diffusion method and performed as recommended by the Clinical and Laboratory Standard Institute (CLSI) M44-A document. Candida isolates investigated produce secondary metabolites gliotoxin with HPLC technique and quantification. Out of 110 Candida isolates, C. albicans (66.36%) was the most frequent isolate, followed by the isolates of C. tropicalis (10.9%) and C. glabrata (6.36%) respectively. Concerning the antifungal susceptibility test, Candida isolates showed a high level of susceptibility to Miconazole (70.9%), Itraconazole (68.2%), and Nystatine (64.5%). The ability of obtained isolates of Candida spp. to produce gliotoxin on RPMI medium was investigated, only 28 isolates had the ability to secret this toxin in culture filtrates. The highest concentrations were detected in C. albicans (1.048 ㎍/ml). Gliotoxin productivity of other Candida species was significantly lower. The retention time for gliotoxin was approximately 5.08 min.

• The Plant Pathology Journal
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• v.33 no.5
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• pp.499-507
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• 2017

In an attempt to develop a biological control agent against mycotoxigenic Fusarium species, we isolated Bacillus amyloliquefaciens strain DA12 from soil and explored its antimicrobial activities. DA12 was active against the growth of mycotoxigenic F. asiaticum, F. graminearum, F. proliferatum, and F. verticillioides both in vitro and in planta (maize). Further screening using dual culture extended the activity range of strain DA12 against other fungal pathogens including Botrytis cinerea, Colletotrichum coccodes, Endothia parasitica, Fusarium oxysporum, Raffaelea quercus-mongolicae, and Rhizoctonia solani. The butanol extract of the culture filtrate of B. amyloliquefaciens DA12 highly inhibited the germination of F. graminearum macroconidia with inhibition rate 83% at a concentration of $31.3{\mu}g/ml$ and 100% at a concentration of $250{\mu}g/ml$. The antifungal metabolite from the butanol extract was identified as iturin A by thin layer chromatography-bioautography. In addition, volatile organic compounds produced by DA12 were able to inhibit mycelial growth of various phytopathogenic fungi. The volatile compounds were identified as 2-heptanone, 5-methyl heptanone and 6-methyl heptanone by gas chromatography-mass spectrometry (GC-MS) analysis. These results indicate that the antagonistic activity of Bacillus amyloliquefaciens DA12 was attributable to iturin A and volatile heptanones, and the strain could be used as a biocontrol agent to reduce the development of Fusarium diseases and mycotoxin contamination of crops.

• Microbiology and Biotechnology Letters
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• v.32 no.1
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• pp.52-59
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• 2004

When both fructose and galactose were added to a production medium as carbon sources, the productivity of PAFS (Psedomonas Antifungal Substance) biosynthesized by Pseudomonas aeruginosa was observed to be reduced significantly due to the well-known phenomenon of catabolite repression. In order to overcome this phenomenon by use of fermentation bioprocess, fed-batch cultivation method was examined. In addition, a high producer mutant strain, AP-20 obtained by a rational screening method was tested for its productivity of PAFS in both batch and fed-batch fermentation processes. Notably fed-batch operation showed approximately 4 fold higher PAFS productivity than traditional batch operation process. It was appeared that galactose was utilized principally for the cell growth of Pseudomonas aeruginosa whereas large portion of fructose was used for the biosynthesis of PAFS. Furthermore it was observed that composition and feeding rate of production media should be optimized even in the fed-batch fermentation bioprocess. As an example, very slow feeding of carbon sources gave rather negative effect on the production of PAFS due to significant limitation of carbon and energy sources available for the producer microorganism.

• Journal of Microbiology and Biotechnology
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• v.28 no.5
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• pp.784-795
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• 2018

Bacillus amyloliquefaciens Pc3 was isolated from Antarctic seawater with antifungal activity. In order to investigate the metabolic regulation mechanism in the biosynthesis of lipopeptides in B. amyloliquefaciens Pc3, GC/MS-based metabolomics was used when exogenous indole was added. The intracellular metabolite profiles showed decreased asparagine, aspartic acid, glutamine, glutamic acid, threonine, valine, isoleucine, hexadecanoic acid, and octadecanoic acid in the indole-treated groups, which were involved in the biosynthesis of lipopeptides. B. amyloliquefaciens Pc3 exhibited a growth promotion, bacterial total protein increase, and lipopeptide biosynthesis inhibition upon the addition of indole. Besides this, real-time PCR analysis further revealed that the transcription of lipopeptide biosynthesis genes ituD, fenA, and srfA-A were downregulated by indole with 22.4-, 21.98-, and 26.0-fold, respectively. It therefore was speculated that as the metabolic flux of most of the amino acids and fatty acids were transferred to the synthesis of proteins and biomass, lipopeptide biosynthesis was weakened owing to the lack of precursor amino acids and fatty acids.

• Journal of Applied Biological Chemistry
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• v.56 no.2
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• pp.109-112
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• 2013

Azole fungicides are one of the most wide-spread antifungal compounds in agriculture and pharmaceutical applications. Their major mode of action is the inhibition of ergosterol biosynthesis, giving depletion of ergosterol, precursors and abnormal steroids. However, metabolic consequences of such inhibition, other than steroidal metabolitesare not well established. Comprehensive metabolic profiles of Saccharomyces cerevisiae has been presented in this study. Wild type yeast was treated either with glucose as control or azole fungicide (ketoconazole). Both polar metabolites and lipids were analyzed with gas chromatography-mass spectrometry. Approximately over 180 metabolites were characterized, among which 18 of them were accumulated or depleted by fungicide treatment. Steroid profile gives the most prominent differences, including the accumulation of lanosterol and the depletion of zymosterol and ergosterol. However, the polar metabolite profile was also highly different in pesticide treatment. The concentration of proline and its precursors, glutamate and ornithine were markedly reduced by ketoconazole. Lysine and glycine level was also decreased while the concentrations of serine and homoserine were increased. The overall metabolic profile indicates that azole fungicide treatment induces the depletion of many polar metabolites, which are important in stress response.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.869-872
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• 2009

The cyclic undecapeptide cyclosporin A (CyA), one of the most valuable immunosuppressive drugs, is produced nonribosomally by a multifunctional cyclosporin synthetase enzyme complex by the filamentous fungus Tolypocladium niveum. To increase CyA productivity by wild-type T. niveum (ATCC 34921), random mutagenesis was first performed using an antifungal agar-plug colony assay (APCA) selection approach. This generated a mutant strain producing more than 9-fold greater CyA than the wild-type strain. Additionally, a foreign bacterial gene, Vitreoscilla hemoglobin gene (VHb), was transformed via protoplast regeneration and its transcription was confirmed by RT-PCR in the UV-irradiated mutant cell. This led to an additional 33.5% increase of CyA production. Although most protoplast-regenerated T. niveum transformants tend to lose CyA productivity, the optimized combination of random mutagenesis and protoplast transformation described here should be an efficient strategy to generate a commercially valuable, yet metabolite low-producing, fungal species, such as CyA-producing T. niveum.

• Journal of Environmental Health Sciences
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• v.31 no.1
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• pp.31-38
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• 2005

An anti-fungal material produced by actinomycetes was isolated from domestic soil. This actinomycetes was identified as Streptomyces albogriseus by 16S rDNA sequence. YEME (yeast extract 4 g, malt extract 10 g, glucose 4 g, D.W 1l, pH 7.00.2) medium was used for production of anti-fungal materials. S. albogriseus was cultured in a shaking incubator for 2 weeks at 150 rpm and $25^{\circ}C$. An anti-fungal material produced by S. albogriseus was identified at 340 nm by uv/vis- spectrometer and it showed powerful anti-fungal activity. This is the first report that secondary metabolite produced by S. albogriseus showed an activity against phytopathogenic fungi such as Collectrichum coccodes, Botrytis cinerea, Cladosporium cucumerinum, Didymella bryoniae.

• The Plant Pathology Journal
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• v.36 no.6
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• pp.570-578
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• 2020

Rhizopus rot is a serious postharvest disease of various crops caused by Rhizopus spp. and controlled mainly by synthetic fungicides. We detected the antifungal activity of a culture extract of Setosphaeria rostrata F3736 against Rhizopus oryzae. The active ingredient was identified as moriniafungin, a known sordarin derivative, which showed minimum inhibitory concentrations of 1-8 ㎍/ml against Colletotrichum spp. and 0.03-0.13 ㎍/ml against Rhizopus spp. in vitro. Moriniafungin showed protective control efficacies against Rhizopus rot on apple and peach fruits. Treatment with 25 ㎍/ml moriniafungin delimited the lesion diameter significantly by 100% on R. oryzae-inoculated apple fruits compared with the non-treated control. Treatment with 0.04 ㎍/ml of moriniafungin reduced the lesion diameter significantly by 56.45%, and treatment with higher concentrations of 0.2-25 ㎍/ml reduced the lesion diameter by 70-90% on Rhizopus stolonifer var. stolonifer-inoculated peach fruit. These results suggest moriniafungin has potential as a control agent of postharvest diseases caused by Rhizopus spp.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.108.1-108
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• 2003

Soil metagenome is untapped total microbial genome including that of the majority of unculturable bacteria present in soil. We constructed soil metagenomic library in Escherichia coli using DNA directly extracted from two different soils, pine tree rhizosphere soil and forest topsoil. Metagenomic libraries constructed from pine tree rhizosphere soil and forest topsoil consisted of approximately 33,700 clones and 112,000 clones with average insert DNA size of 35-kb, respectively. Subsequently, we screened the libraries to select clones with antimicrobial activities against Saccharomyces cerevisiae and Agrobacterium tumefaciens using double agar layer method. So far, we have a clone active against S. cerevisiae and a clone active against A. tumefaciens from the forest topsoil library. In vitro mutagenesis and DNA sequence analysis of the antifungal clone revealed the genes involved in the biosynthesis of antimicrobial secondary metabolite. Metagenomic libraries constructed in this study would be subject to search for diverse genetic resources related with useful microbial products.