• Title/Summary/Keyword: Dual culture assay

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Biocontrol of gray mold of cucumber and tomato by epihpytic bacteria in field condition

  • Lee, Sang-Yeob;Lee, Sang-Bum;Kim, Il-Yon;Kim, Yong-Ki
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.104.2-105
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    • 2003
  • Bacterial isolates, CC178, PTC25, HC39 and KY165 originally obtained from the leaves of cucumber or tomato were selected for biocontrol agents against gray mold of cucumber and tomato by in vivo cucumber seedling assay. Each suspension of the selected epiphytic bacteria were sprayed three times at seven-day interval from early stage of cucumber in a field. Incidence of gray mold on cucumber fruits treated with isolates CC178, PTC25, HC39 and KY165 was 15.3%, 18.2%, 23.6%, and 10.4%, respectively, whereas that of control was 38.0% after 7 days of final spray. On the other hand, treatment with the selected isolates, CC178, PTC25, HC39, and KY165 on tomato showed 2.2%, 1.3%, 2.9%, and 3.5% in the incidence of gray mold on leaves, whereas that of control was 9.3%. All selected isolates had strong antagonistic activity against Botrytis cinerea on dual culture plate assay.

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Antifungical Activity of Autochthonous Bacillus subtilis Isolated from Prosopis juliflora against Phytopathogenic Fungi

  • Abdelmoteleb, Ali;Troncoso-Rojas, Rosalba;Gonzalez-Soto, Tania;Gonzalez-Mendoza, Daniel
    • Mycobiology
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    • v.45 no.4
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    • pp.385-391
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    • 2017
  • The ability of Bacillus subtilis, strain ALICA to produce three mycolytic enzymes (chitinase, ${\beta}$-1,3-glucanase, and protease), was carried out by the chemical standard methods. Bacillus subtilis ALICA was screened based on their antifungal activity in dual plate assay and cell-free culture filtrate (25%) against five different phytopathogenic fungi Alternaria alternata, Macrophomina sp., Colletotrichum gloeosporioides, Botrytis cinerea, and Sclerotium rolfesii. The B. subtilis ALICA detected positive for chitinase, ${\beta}$-1,3-glucanase and protease enzymes. Fungal growth inhibition by both strain ALICA and its cell-free culture filtrate ranged from 51.36% to 86.3% and 38.43% to 68.6%, respectively. Moreover, hyphal morphological changes like damage, broken, swelling, distortions abnormal morphology were observed. Genes expression of protease, ${\beta}$-1,3-glucanase, and lipopeptides (subtilosin and subtilisin) were confirmed their presence in the supernatant of strain ALICA. Our findings indicated that strain ALICA provided a broad spectrum of antifungal activities against various phytopathogenic fungi and may be a potential effective alternative to chemical fungicides.

Antifungal Activity of Lactobacillus plantarum Isolated from Kimchi (김치로부터 항진균 활성 Lactobacillus plantarum의 분리 및 특성 규명)

  • Yang, Eun-Ju;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.276-284
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    • 2008
  • A lactic acid bacterium having antifungal activity was isolated from kimchi. It was identified as Lactobacillus plantarum based on its morphological and biochemical properties, and 16S rRNA sequence, and designated as Lb. plantarum AF1. This isolate inhibited the growth of Aspergillus flavus ATCC 22546, A. fumigatus ATCC 96918, A. petrakii PF-1, A. ochraceus PF-2, A. nidulans PF-3, Epicoccum nigrum KF-1, and Cladosporium gossypiicola KF-2 under a dual culture overlay assay. Also, the antimicrobial activity was found to be active against various species of Gram-positive and Gram-negative bacteria. The antifungal activity was found to be stable after heat ($121^{\circ}C$, 15 min) and proteolytic enzyme treatment, but it was unstable over pH 5.0. The antifungal compound(s) was estimated to have a low molecular mass (below 3,000 Da).

Screening and Evaluation of Streptomyces Species as a Potential Biocontrol Agent against a Wood Decay Fungus, Gloeophyllum trabeum

  • Jung, Su Jung;Kim, Nam Kyu;Lee, Dong-Hyeon;Hong, Soon Il;Lee, Jong Kyu
    • Mycobiology
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    • v.46 no.2
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    • pp.138-146
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    • 2018
  • Two-hundred and fifty-five strains of actinomycetes isolated from soil samples were screened for their antagonistic activities against four well-known wood decay fungi (WDF), including a brown rot fungus, Gloeophyllum trabeum and three white rot fungi Donkioporia expansa, Trametes versicolor, and Schizophyllum commune. A dual culture assay using culture media supplemented with heated or unheated culture filtrates of selected bacterial strains was used for the detection of their antimicrobial activity against four WDF. It was shown that Streptomyces atratus, S. tsukiyonensis, and Streptomyces sp. greatly inhibited the mycelial growth of the WDF tested compared with the control. To evaluate the biocontrol efficacy of S. atratus, S. tsukiyonensis, and Streptomyces sp., wood blocks of Pinus densiflora inoculated with three selected Streptomyces isolates were tested for weight loss, compression strength (perpendicular or parallel to the grain), bending strength, and chemical component changes. Of these three isolates used, Streptomyces sp. exhibited higher inhibitory activity against WDF, especially G. trabeum, as observed in mechanical and chemical change analyses. Scanning electron microscopy showed that cell walls of the wood block treated with Streptomyces strains were thicker and collapsed to a lesser extent than those of the non-treated control. Taken together, our findings indicate that Streptomyces sp. exhibits the potential to be used as a biocontrol agent for wood decay brown rot fungus that causes severe damage to coniferous woods.

Selection of Antagonistic Soil Actinomycetes Against Both Colletotrichum orbiculare and Botrytis cinerea in Cucumber Plants (오이 탄저병원균과 잿빛곰팡이병원균에 억제 효과가 있는 방선균 선발)

  • Kwak, Hwa-Sook;Kim, Jiwon;Park, Jin Woo;Sang, Mee Kyung
    • Korean Journal of Organic Agriculture
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    • v.29 no.4
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    • pp.575-588
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    • 2021
  • The purpose of this study is to select actinomycetes with antifungal activity against Botrytis cinerea and Colletotrichum orbiculare, which are airborne pathogens in cucumber plants. In 2020, 560 species of actinomycetes were isolated from rhizome soils of various vegetables in Korea; 7 strains showing simultaneous antifungal activity against two pathogens were selected. Finally, strain S20-465 was selected through dual culture and plant assay. This strain was identified as Streptomyces sp. based on 16S rRNA analysis. The culture filtrate of strain S20-465 inhibited mycelial growth of both pathogens by more than 60%. In addition, when cucumber plants were treated with 20-fold and 40-fold diluted culture filtrates of S20-465, lesions caused by B. cinerea and C. orbiculare on cucumber leaves were significantly reduced compared to the control. This results suggest that strain S20-465 produces specific secondary metabolites with antifungal activity against both pathogens.

Identification and Characterization of Trichoderma Species Damaging Shiitake Mushroom Bed-Logs Infested by Camptomyia Pest

  • Kim, Jun Young;Kwon, Hyuk Woo;Yun, Yeo Hong;Kim, Seong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.26 no.5
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    • pp.909-917
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    • 2016
  • The shiitake mushroom industry has suffered from Camptomyia (gall midges) pest, which feeds on the mycelium of shiitake mushroom during its cultivation. It has been postulated that fungal damage of shiitake bed-logs is associated with infestation by the insect pest, but this is not well understood. To understand the fungal damage associated with Camptomyia pest, various Trichoderma species were isolated, identified, and characterized. In addition to two previously known Trichoderma species, T. citrinoviride and T. deliquescens, two other Trichoderma species, T. harzianum and T. atroviride, were newly identified from the pestinfested bed-log samples obtained at three mushroom farms in Cheonan, Korea. Among these four species, T. harzianum was the most evident. The results of a chromogenic media-based assay for extracellular enzymes showed that these four species have the ability to produce amylase, carboxyl-methyl cellulase, avicelase, pectinase, and ß-glucosidase, thus indicating that they can degrade wood components. A dual culture assay on PDA indicated that T. harzianum, T. atroviride, and T. citrinoviride were antagonistic against the mycelial growth of a shiitake strain (Lentinula edodes). Inoculation tests on shiitake bed-logs revealed that all four species were able to damage the wood of bed-logs. Our results provide evidence that the four green mold species are the causal agents involved in fungal damage of shiitake bed-logs infested by Camptomyia pest.

An Interferon Resistance Induced by the Interaction between HCV NS5B and Host p48 (C형 간염 바이러스 NS5B 단백질과 숙주의 p48 단백질의 상호작용에 의한 인터페론 저항성의 유도)

  • Park, So-Yeon;Lee, Jong-Ho;Myung, Hee-Joon
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.353-359
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    • 2008
  • Hepatitis C virus (HCV) is known as the causative agent of blood transmitted hepatitis. Two viral proteins, E2 and NS5A, are known to exert interferon resistance of HCV via PKR pathway. Here, we report a third protein, the RNA-dependent RNA polymerase (NS5B) of HCV, induced interferon resistance inhibiting p56 pathway. p56 was shown to interact with p48 subunit of eukaryotic initiation factor 3 (eIF3). This interaction inhibited formation of ternary complex in translation initiation. Using dual reporter assay system, we observed that the translation decreased when interferon alpha was added to the culture. But, in the presence of HCV NS5B, the translation partly recovered. NS5B and p48 subunit of eIF3 were shown to interact. This interaction seems to inhibit the interaction between p48 and p56. This is the first report that a virus exerts interferon resistance via p56 pathway.

Antagonistic Potential of Fluorescent Pseudomonads and Control of Crown and Root Rot of Cucumber Caused by Phythophtora drechsleri

  • Shirzad, Akbar;Fallahzadeh-Mamaghani, Vahid;Pazhouhandeh, Maghsoud
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.1-9
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    • 2012
  • In this study, 200 isolates of fluorescent pseudomonads were isolated from different fields of East and West Azarbaijan and Ardebil provinces of Iran. These bacterial isolates were screened on the basis of a dual culture assay, the presence of known antibiotic genes, and their ability to successfully colonize roots and to promote plant growth. Twelve isolates exhibited 30% or more inhibition of mycelia growth of $P.$ $drechsleri$. Genes encoding production of the antibiotics 2,4-diacetylphloroglucinol, phenazine-1-carboxylic acid, and pyoluteorin were detected in some strains but none of the strains possessed the coding gene for production of antibiotic pyrrolnitrin. In an $in$ $vitro$ test for root colonization, the population density on roots of plants treated with most of the above strains was more than 6 $\log_{10}$ CFU $g^{-1}$ roots, with a maximum of 7.99 $\log_{10}$ CFU $g^{-1}$ roots for strain 58A. Most of the strains promoted significant plant growth in comparison to non-treated controls. In green house studies, the percentage of healthy plants in pots treated with strains 58A and 8B was 90.8% and 88.7%, respectively. The difference between these treatments and treatment with the fungicide metalaxyl was not significant.

Chemical and Biological Controls of Balloon Flower Stem Rots Caused by Rhizoctonia solani and Sclerotinia sclerotiorum

  • Lee, Young-Hee;Cho, Young-Son;Lee, Shin-Woo;Hong, Jeum-Kyu
    • The Plant Pathology Journal
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    • v.28 no.2
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    • pp.156-163
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    • 2012
  • Stem rots caused by Rhizoctonia solani and Sclerotinia sclerotiorum have been known as devastating diseases in balloon flower plants. Antifungal activities of four fungicides, azoxystrobin, polyoxin B, trifloxystrobin and validamycin A were evaluated in vitro, showing effective suppression with mycelial growth of the fungal isolates on PDA media. Efficacies of the four fungicides were also demonstrated in stem tissues of balloon flower plants against R. solani and S. sclerotiorum. A commercially available Bacillus subtilis strain Y1336 was tested in terms of antagonistic biological control of stem rot disease of balloon flower plants. The bacterial strain revealed its antifungal activities against R. solani and S. sclerotiorum demonstrated by dual culture tests using paper discs and two plant pathogenic fungi on PDA media, as well as by plant inoculation assay, indicating that this antagonistic bacterial strain can be incorporated into disease management program for balloon flower stem rot diseases together with the four chemical fungicides.

Antagonistic Activity of Siderophore-Producing Bacteria from Black Rice Rhizosphere against Rice Blast Fungus Pyricularia oryzae

  • Nabila, Nabila;Kasiamdari, Rina Sri
    • Microbiology and Biotechnology Letters
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    • v.49 no.2
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    • pp.217-224
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    • 2021
  • Rice blast caused by Pyricularia oryzae, which is a major threat to food security worldwide, markedly decreases the yield of rice. Some rhizobacteria called 'plant growth-promoting rhizobacteria' inhibit plant pathogens and improve plant growth by secreting iron-chelating siderophores. The decreased availability of iron adversely affects the survival of pathogens, especially fungal pathogens, in the rhizosphere. This study aimed to determine the morphological diversity of siderophore-producing bacteria, analyze the type of siderophores produced by the bacteria, and examine their growth-inhibitory activity against Pyricularia oryzae. The rhizobacteria were isolated from the rhizosphere of Sembada Hitam variety of black rice plants in Pakem, Sleman, Yogyakarta, Indonesia. In total, 12 distinct isolates were screened for the production of siderophores. It was found that 9 out of 12 bacteria produced siderophore and most of them were Gram positive bacteria. The best siderophore-producing isolates with different type of siderophore were used in further studies. The IS3 and IS14 isolates were found to be the best siderophore producer that produced hydroxamate and mixed type of hydroxamate-carboxylate type of siderophore, respectively. In the dual culture assay, IS14 showed a strong antagonistic effect against Pyricularia oryzae by the 81.17% inhibition.