• Mycobiology
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• v.48 no.1
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• pp.37-43
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• 2020

A serious leaf disease caused by Colletotrichum dematium was found during the cultivation of Sarcandra glabra in Jingxi, Rong'an, and Donglan Counties in Guangxi Province, which inflicted huge losses to plant productivity. Biological control gradually became an effective control method for plant pathogens. Many studies showed that the application of actinomycetes in biological control has been effective. Therefore, it may be of great significance to study the application of actinomycetes on controlling the diseases caused by S. glabra. Strains of antifungal actinomycetes capable of inhibiting C. dematium were identified, isolated and screened from healthy plants tissues and the rhizospheres in soils containing S. glabra. In this study, 15 actinomycetes strains were isolated and among these, strains JT-2F, DT-3F, and JJ-3F, appeared to show antagonistic effects against anthracnose of S. glabra. The strains JT-2F and DT-3F were isolated from soil, while JJ-3F was isolated from plant stems. The antagonism rate of strain JT-2F was 86.75%, which was the highest value among the three strains. Additionally, the JT-2F strain also had the strongest antagonistic activity when the antagonistic activities were tested against seven plant pathogens. Strain JT-2F is able to produce proteases and cellulase to degrade the protein and cellulose components of cell walls of C. dematium, respectively. This results in mycelia damage which leads to inhibition of the growth of C. dematium. Strain JT-2F was identified as Streptomyces tsukiyonensis based on morphological traits and 16S rDNA sequence analysis.

• Microbiology and Biotechnology Letters
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• v.31 no.4
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• pp.421-428
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• 2003

To develop the functional-compost containing antifungal substance by using antagonistic microorganisms, Spinacia oleracea L and Rhizoctonia solani Kuhn O-28 were used as a model plant and phytopathogen, respectively. Total 80 strains were isolated from the compost of various waste foods mixture processed for a year. Among them, No.15-2 strain was selected due to its highest antifungal activity against R. solani Kuhn O-28 and was identified phyno- and phylogenotypically as Burkholderia cepacia genomovar V. which is rare probability in pathogen, by 16S rDNA sequencing and specific primer pair PCR method. B. cepacia No.15-2 preferentially dominated during the compost and its cell numbers were maintained almost $${\times}$10^{13}$ cuf/g for 15 days. The morbidity caused by R. solani Kuhn O-28 in S. oleracea L cultivation was reduced to 40% by addition of B. cepacia No.15-2. In conclusion, the antifungal compost using B. cepacia No.15-2 could be applied to biocontrol of various crops blights caused by fungal pathogen.

• Journal of Microbiology and Biotechnology
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• v.24 no.9
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• pp.1216-1225
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• 2014

Biofilm-related infections of Candida albicans are a frequent cause of morbidity and mortality in hospitalized patients, especially those with immunocompromised status. Options of the antifungal drugs available for successful treatment of drug-resistant biofilms are very few, and as such, new strategies need to be explored against them. The aim of this study was to evaluate the efficacy of phenylpropanoids of plant origin against planktonic cells, important virulence factors, and biofilm forms of C. albicans. Standard susceptibility testing protocol was used to evaluate the activities of 13 phenylpropanoids against planktonic growth. Their effects on adhesion and yeast-to-hyphae morphogenesis were studied in microplate-based methodologies. An in vitro biofilm model analyzed the phenylpropanoid-mediated prevention of biofilm development and mature biofilms using XTT-metabolic assay, crystal violet assay, and light microscopy. Six molecules exhibited fungistatic activity at ${\leq}0.5mg/ml$, of which four were fungicidal at low concentrations. Seven phenylpropanoids inhibited yeast-to-hyphae transition at low concentrations (0.031-0.5 mg/ml), whereas adhesion to the solid substrate was prevented in the range of 0.5-2 mg/ml. Treatment with ${\leq}0.5mg/ml$ concentrations of at least six small molecules resulted in significant (p < 0.05) inhibition of biofilm formation by C. albicans. Mature biofilms that are highly resistant to antifungal drugs were susceptible to low concentrations of 4 of the 13 molecules. This study revealed phenylpropanoids of plant origin as promising candidates to devise preventive strategies against drug-resistant biofilms of C. albicans.

• Journal of Microbiology and Biotechnology
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• v.30 no.7
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• pp.1018-1026
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• 2020

Rosemary essential oil was evaluated for antifungal potentiality against six major ginseng pathogens: Sclerotinia sclerotiorum, Sclerotinia nivalis, Cylindrocarpon destructans, Alternaria panax, Botrytis cinerea, and Fusarium oxysporum. The in vitro fungicidal effects of two commonly used fungicides, namely mancozeb and fenhexamid, and the volatile organic compounds (VOCs) of Trichoderma koningiopsis T-403 on the mycelial growth were investigated. The results showed that rosemary essential oil is active against all of the pathogenic strains of ginseng root rot, whereas rosemary oil displayed high ability to inhibit the Sclerotinia spp. growth. The highest sensitivity was S. nivalis, with complete inhibition of growth at 0.1% v/v of rosemary oil, followed by Alternaria panax, which exhibited 100% inhibition at 0.3% v/v of the oil. Minimum inhibitory concentrations (MICs) of rosemary oil ranged from 0.1 % to 0.5 % (v/v). Chemical analysis using GC-MS showed the presence of thirty-two constituents within rosemary oil from R. officinals L. Camphore type is the most frequent sesquiterpene in rosemary oil composition. Mancozeb and fenhexamid showed their highest inhibition effect (45% and 30%, respectively) against A. panax. T. koningiopsis T-403 showed its highest inhibition effect (84%) against C. destructans isolate. This study may expedite the application of antifungal natural substances from rosemary and Trichoderma in the prevention and control of phytopathogenic strains in ginseng root infections.

• The Plant Pathology Journal
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• v.32 no.1
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• pp.16-24
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• 2016

In this study, antifungal activity of essential oils of Cymbopogon citratus and Ocimum basilicum and two fungicides Mancozeb and Metalaxyl-Mancozeb in six different concentrations were investigated for controlling three species of Phytophthora, including P. capsici, P. drechsleri and P. melonis on pepper, cucumber and melon under in vitro and greenhouse conditions, respectively. Under the in vitro condition, the median effective concen- tration ($EC_{50}$) values (ppm) of plant essential oils and fungicides were measured. In greenhouse, soil infested with Phytophthora species was treated by adding 50 ml of essential oils and fungicides (100 ppm). Disease severity was determined after 28 days. Among two tested plant essential oils, C. citratus had the lowest $EC_{50}$ values for inhibition of the mycelial growth of P. capsici (31.473), P. melonis (33.097) and P. drechsleri (69.112), respectively. The mean $EC_{50}$ values for Metalaxyl-Mancozeb on these pathogens were 20.87, 20.06 and 17.70, respectively. Chemical analysis of plant essential oils by GC-MS showed that, among 42 compounds identified from C. citratus, two compounds ${\beta}$-geranial (${\alpha}$-citral) (39.16%) and z-citral (30.95%) were the most abundant. Under the greenhouse condition, Metalaxyl-Mancozeb caused the greatest reduction in disease severity, 84.2%, 86.8% and 92.1% on melon, cucumber, and pepper, respectively. The C. citratus essential oil reduced disease severity from 47.4% to 60.5% compared to the untreated control ($p{\leq}0.05$). Essential oils of O. basilicum had the lowest effects on the pathogens under in vitro and greenhouse conditions. These results show that essential oils may contribute to the development of new antifungal agents to protect the crops from Phytophthora diseases.

• Research in Plant Disease
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• v.16 no.2
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• pp.170-175
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• 2010

A monoterpenoid benzylideneacetone (BZA) is a bacterial metabolite isolated from culture broth of an entomopathogenic bacterium, Xenorhabdus nematophila K1. It was tested in this study the control efficacy of the metabolite against two major fungal diseases occurring in red-pepper plants. BZA exhibited significant antifungal activities against Phytophthora capsici and Colletotrichum acutatum. Under natural light conditions, the antifungal activity of BZA was maintained for more than sixty days. The antifungal activity of BZA was not lost even in soil because the incidence of Phytophthora blight against red-pepper plants was significantly reduced when the suspensions of P. capsici were poured to the rhizosphere soils mixed with BZA. Application of the BZA suspension spray to the fruit surface infected with C. acutatum significantly suppressed the disease occurrence of anthracnose on the red-pepper plants. These results suggest that BZA can be used to develop a promising agrochemical to control phytophthora blight and anthracnose of redpepper plants.

• Biomolecules & Therapeutics
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• v.26 no.5
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• pp.494-502
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• 2018

Breast cancer is currently the most prevalent cancer in women, and its incidence increases every year. Azole antifungal drugs were recently found to have antitumor efficacy in several cancer types. They contain an imidazole (clotrimazole and ketoconazole) or a triazole (fluconazole and itraconazole) ring. Using human breast adenocarcinoma cells (MCF-7 and MDA-MB-231), we evaluated the effects of azole drugs on cell proliferation, apoptosis, cell cycle, migration, and invasion, and investigated the underlying mechanisms. Clotrimazole and ketoconazole inhibited the proliferation of both cell lines while fluconazole and itraconazole did not. In addition, clotrimazole and ketoconazole inhibited the motility of MDA-MB-231 cells and induced $G_1$-phase arrest in MCF-7 and MDA-MB-231 cells, as determined by cell cycle analysis and immunoblot data. Moreover, Transwell invasion and gelatin zymography assays revealed that clotrimazole and ketoconazole suppressed invasiveness through the inhibition of matrix metalloproteinase 9 in MDA-MB-231 cells, although no significant changes in invasiveness were observed in MCF-7 cells. There were no significant changes in any of the observed parameters with fluconazole or itraconazole treatment in either breast cancer cell line. Taken together, imidazole antifungal drugs showed strong antitumor activity in breast cancer cells through induction of apoptosis and $G_1$ arrest in both MCF-7 and MDA-MB-231 cells and suppression of invasiveness via matrix metalloproteinase 9 inhibition in MDA-MB-231 cells. Imidazole drugs have well-established pharmacokinetic profiles and known toxicity, which can make these generic drugs strong candidates for repositioning as antitumor therapies.

• Journal of Ginseng Research
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• v.42 no.3
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• pp.304-311
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• 2018

Background: Ginseng black spot disease resulting from Alternaria panax Whuetz is a common soil-borne disease, with an annual incidence rate higher than 20-30%. In this study, the bacterial strains with good antagonistic effect against A. panax are screened. Methods: A total of 285 bacterial strains isolated from ginseng rhizosphere soils were screened using the Kirby-Bauer disk diffusion method and the Oxford cup plate assay. We analyzed the antifungal spectrum of SZ-22 by confronting incubation. To evaluate the efficacy of biocontrol against ginseng black spot and for growth promotion by SZ-22, we performed pot experiments in a plastic greenhouse. Taxonomic position of SZ-22 was identified using morphology, physiological, and biochemical characteristics, 16S ribosomal DNA, and gyrB sequences. Results: SZ-22 (which was identified as Brevundimonas terrae) showed the strongest inhibition rate against A. panax, which showed 83.70% inhibition, and it also provided broad-spectrum antifungal effects. The inhibition efficacies of the SZ-22 bacterial suspension against ginseng black spot reached 82.47% inhibition, which is significantly higher than that of the 25% suspension concentrate azoxystrobin fungicide treatment (p < 0.05). Moreover, the SZ-22 bacterial suspension also caused ginseng plant growth promotion as well as root enhancement. Conclusion: Although the results of the outdoor pot-culture method were influenced by the pathogen inoculum density, the cropping history of the field site, and the weather conditions, B. terrae SZ-22 controlled ginseng black spot and promoted ginseng growth successfully. This study provides resource for the biocontrol of ginseng black spot.

• Journal of Microbiology and Biotechnology
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• v.7 no.2
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• pp.107-113
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• 1997

A strain producing a high amount of chitinase was isolated from soil, identified as Pseudomonas sp., and tentatively named Pseudomonas sp. YHS-A2. An extracellular chitinase of Pseudomonas sp. YHS-A2 was purified according to the procedure of ammonium sulfate saturation, affinity adsorption, Sephadex G-100 gel filtration and Phenyl-sepharose CL-4B hydrophobic interaction column chromatography. The molecular weight of the purified enzyme was estimated to be 55 kDa on SDS-PAGE was confirmed by active staining. Optimal pH and temperature of the enzyme are pH 7.0 and $50^{\circ}C$, respectively, and the enzyme is stable between pH 5.0 and 8.0 and below $50^{\circ}C$. The main products of colloidal chitin by the chitinase were N-acetyl-D-glucosamine and N,N'-diacetylchitobiose both of which were detected by HPLC analysis. The enzyme is supposed to be a random-type endochitinase which can degrade any position of ${\beta}$-l,4-linkages of chitin and chitooligosaccharides. The chitinase inhibited the growth of some phytopathogenic fungi, Fusarium oxysporum, Botrytis cineria, and Mucor rouxii and these antifungal effects were thought to be due to the characteristics of endochitinase.

• Journal of Microbiology and Biotechnology
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• v.29 no.1
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• pp.66-78
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• 2019

In this study, strain KNU17Pc1 was tested for its antifungal activity against Rhizoctonia solani AG-1(IA), which causes banded leaf and sheath blight (BLSB) of maize. KNU17Pc1 was tested further for its broad-spectrum antifungal activity and in vitro plant growth promoting (PGP) traits. In addition, the in vivo effects of KNU17Pc1 on reduction of BLSB severity and seedling growth promotion of two maize cultivars under greenhouse conditions were investigated. On the basis of multilocus sequence analysis (MLSA), KNU17Pc1 was confirmed as P. chlororaphis subsp. aurantiaca. The study revealed that KNU17Pc1 had strong in vitro antifungal activity and was effective toward all in vitro PGP traits except phosphate solubilization. In this study, for the first time, a strain of P. chlororaphis against Colletotrichum dematium, Colletotrichum gloeosporioides, Fusarium oxysporum f.sp. melonis, Fusarium subglutinans and Stemphylium lycopersici has been reported. Further biochemical studies showed that KNU17Pc1 was able to produce both types of phenazine derivatives, PCA and 2-OH-PCA. In addition, solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) analysis identified 13 volatile organic compounds (VOCs) in the TSB culture of KNU17Pc1, 1-undecene being the most abundant volatile. Moreover, for the first time, Octamethylcyclotetrasiloxan (D4), dimethyl disulfide, 2-methyl-1,3-butadiene and 1-undecene were detected in P. chlororaphis. Furthermore, this study reported for the first time the effectiveness of P. chlororaphis to control BLSB of maize. Hence, further studies are necessary to test the effectiveness of KNU17Pc1 under different environmental conditions so that it can be exploited further for biocontrol and plant growth promotion.