• 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.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.104.1-104
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• 2003

To screen for potential biocontrol agents against postharvest disease of garlics caused by Penicillium hirsutum, a total of 933 isolates (432 fungi and 501 bacteria) were isolated from the rhizoshere or rhizoplane of garlics. Among them, Pantoea agglomerans isolate 59-4 (Pa 59-4) was selected for a potential biocontrol agent by in vivo wounded garlic bulb assay, When the spore suspension (10$\^$5/ spores/$m\ell$) of Penicillium hirsutum was co-inoculated with spore or cell suspension of each fungal or bacterial isolate on wounded garlics, the isolate highly suppressed disease development. Soaking garlic bulbs in the suspension of Pa 59-4 significantly reduced garlic decay from p. hirsutum. However, Pa 59-4 did not inhibit the mycelial growth of P. hirsutum in dual-culture with P. hirsutum on Tryptic soy agar. In order to elucidate mode of action of Pa 59-4 nutrient competition between Pa 59-4 and P. hirsutum was investigated using tissue culture plates with cylinder inserts containing defusing membrane reported by Janisiewicz et al. The results showed that Pa 59-4 effectively suppressed spore germination and mycelial growth of blue mold in the low concentration (0.5％) of garlic juice, but did not suppress those of blue mold in the higher concentration (5％) of garlic juice. This result suggests that the mechanism in biocontrol of garlic blue mold by Pa 59-4 may involve in nutrient competition with P. hirsutum on garlic bulbs.

• The Plant Pathology Journal
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• v.24 no.4
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• pp.453-460
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• 2008

In the course of in vitro and in vivo screening for bioherbicidal agents, a hyphomycete fungus, Myrothecium sp. F0252 was selected as a candidate for the biocontrol of weeds. The isolate was identified as Myrothecium roridum Tode ex. Fries based on the morphological characteristics and 18S ribosomal DNA sequence analysis and registered as Myrothecium roridum F0252. In order to evaluate the in vitro effect of M. roridum F0252 on germination of ladino clover and white clover (Trifolium repens L.) seeds, spore solution of the fungus was employed in two concentrations, $6.5{\times}10^6$ and $2.5{\times}10^7$ spores per mL and then inoculated to the seeds. The fungal spores inhibited the seed germination, infected the seedlings, and caused an abnormal withering and inhibition of seedling growth. In addition, when the herbicidal activity of crude ethyl acetate extract from the liquid culture was assessed on a mini-plant, duck-weed (Lemna paucicostata (L.) Hegelm.), the extract showed high inhibitory effect at the level of $12.5{\mu}g$ per mL. On the other hand, in vivo herbicidal activity of M. roridum F0252 was evaluated by a whole plant spray method. M. roridum F0252 exhibited strong and broad-spectrum herbicidal activity. The herbicidal values ranged from 95-100% against 7 weeds, including Abutilon avicennae and Xanthium strumarium, and 70-80% against Digitaria sanguinalis and Sagittaria pygmaea. When the nutritional utilization (95 carbon sources) pattern of M. roridum F0252 was investigated, it varied with water activity ($a_w$) and temperature conditions, supplying good, basic information in regard to nutritional utilization for proper cultivation and formulation. Our results showed that M. roridum F0252 might be used as a potential biocontrol agent against weedy plants.

• Korean Journal of Soil Science and Fertilizer
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• v.49 no.4
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• pp.393-400
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• 2016

Fusarium graminearum is the main cause of substantial economic loss in wheat production. The aim of this study is to investigate biocontrol potential of Lysobacter antibioticus HS124 against F. graminearum and to purify an antifungal compound. In preliminary study, n-butanol crude extract revealed destructive alterations in the hyphal morphology of F. graminearum and almost degraded with $1,000{\mu}g\;mL^{-1}$ concentration. For further study, the antifungal compound extracted from the n-butanol crude extract of L. antibioticus HS124 was identified as N-Butyl-tetrahydro-5-oxofuran-2-carboxamide ($C_9H_{16}NO_3$) using NMR ($^1H-NMR$, $^{13}C-NMR$, $^1H-^1H\;COSY$, HMBC, and HMQC), and HR-ESI-MS analysis. To our knowledge, N-Butyl-tetrahydro-5-oxofuran-2-carboxamide may be a novel compound with molecular weight of 186.1130. The minimum inhibitory concentration value of antifungal compound was $62.5{\mu}g\;mL^{-1}$ against F. graminearum. In an in vivo pot experiment, crown rot disease from F. graminearum was inhibited when wheat seeds were treated with both HS124 culture and F. graminearum. Growth of wheat seedling was enhanced by treatment of HS124 compared to control. Our results suggest that L. antibioticus HS124 characterized in this study could be successfully used to control F. graminearum and could be used as an alternative to chemical fungicides in modern agriculture.

• The Plant Pathology Journal
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• v.34 no.3
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• pp.208-217
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• 2018

In this study, samples were collected from the leaves and stems of healthy wild Pistachio trees (Pistacia atlantica L.) from various locations of Baneh and Marivan regions, Iran. In total, 61 endophytic bacteria were isolated and grouped according to phenotypic properties. Ten selected isolates from each group were further identified by partial sequencing of the 16S rRNA gene. Based on the results, isolates were identified as bacteria belonging to Pseudomonas, Stenotrophomonas, Bacillus, Pantoea and Serratia genus. The ability of these isolates was evaluated to phytohormone production such as auxin and gibberellin, siderophore production, phosphate solubilization, atmospheric nitrogen fixation, protease and hydrogen cyanide production. All strains were able to produce the plant growth hormone auxin and gibberellin in different amounts. The majority of strains were able to solubilize phosphate. The results of atmospheric nitrogen fixation ability, protease and siderophore production were varied among strains. Only Ba66 could produce a low amount of hydrogen cyanide. The results of biocontrol assay showed that Pb78 and Sp15 strains had the highest and lowest inhibition effects on bacterial plant pathogens, Pseudomonas syringae pv. syringae Pss20 and Pseudomonas tolaasii Pt18 under in vitro condition. Pb3, Pb24 and Pb71 strains significantly promote root formation on carrot slices. To our knowledge this is the first report of the isolation of endophytic bacterial strains belonging to Pantoea, Bacillus, Pseudomonas, Serratia and Stenotrophomonas genus from wild pistachio trees with plant growth promoting potential and biocontrol activity.

• The Plant Pathology Journal
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• v.34 no.2
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• pp.126-138
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• 2018

Rice blast caused by Magnaporthe oryzae is a major disease. In the present study, we aimed to identify and evaluate the novel bacterial isolates from rice rhizosphere for biocontrol of M. oryzae pathogen. Sixty bacterial strains from the rice plant's rhizosphere were tested for their biocontrol activity against M. oryzae under in vitro and in vivo. Among them, B. amyloliquefaciens had significant high activity against the pathogen. The least disease severity and highest germination were recorded in seeds treated with B. amyloliquefaciens UASBR9 (0.96 and 98.00%) compared to untreated control (3.43 and 95.00%, respectively) under in vivo condition. These isolates had high activity of enzymes in relation to growth promoting activity upon challenge inoculation of the pathogen. The potential strains were identified based on 16S rRNA gene sequencing and dominance of these particular genes were associated in Bacillus strains. These strains were also confirmed for the presence of antimicrobial peptide biosynthetic genes viz., srfAA (surfactin), fenD (fengycin), spaS (subtilin), and ituC (iturin) related to secondary metabolite production (e.g., AMPs). Overall, the results suggested that application of potential bacterial strains like B. amyloliquefaciens UASBR9 not only helps in control of the biological suppression of one of the most devastating rice pathogens, M. grisea but also increases plant growth along with a reduction in application of toxic chemical pesticides.

• Journal of Microbiology and Biotechnology
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• v.27 no.6
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• pp.1157-1162
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• 2017

The goal of this study was to identify and characterize selected Trichoderma isolates by metabolic profiling and enzyme assay for evaluation of their potential as biocontrol agents against plant pathogens. Trichoderma isolates were obtained from the Rural Development Administration Genebank Information Center (Wanju, Republic of Korea). Eleven Trichoderma isolates were re-identified using ribosomal DNA internal transcribed spacer (ITS) regions. ITS sequence results showed new identification of Trichoderma isolates. In addition, metabolic profiling of the ethyl acetate extracts of the liquid cultures of five Trichoderma isolates that showed the best anti-Phytophthora activities was conducted using gas chromatography-mass spectrometry. Metabolic profiling revealed that Trichoderma isolates shared common metabolites with well-known antifungal activities. Enzyme assays indicated strong cell wall-degrading enzyme activities of Trichoderma isolates. Overall, our results indicated that the selected Trichoderma isolates have great potential for use as biocontrol agents against plant pathogens.

• Mycobiology
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• v.49 no.1
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• pp.69-77
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• 2021

The application of antagonistic fungi for plant protection has attracted considerable interest because they may potentially replace the use of chemical pesticides. Antipathogenic activities confirmed in volatile organic compounds (VOCs) from microorganisms have potential to serve as biocontrol agents against pre- and post-harvest diseases. In the present study, we investigated Galactomyces fungi isolated from rotten leaves and the rhizosphere of cherry tomato (Lycopersicon esculentum var. cerasiforme). VOCs produced by Galactomyces fungi negatively affected the growth of phytopathogenic fungi and the survival of nematodes. Mycelial growths of all nine examined phytopathogenic fungi were inhibited on agar plate, although the inhibition was more intense in Athelia rolfsii JYC2163 and Cladosporium cladosporioides JYC2144 and relatively moderate in Fusarium sp. JYC2145. VOCs also efficiently suppressed the spore germination and mycelial growth of A. rolfsii JYC2163 on tomatoes. The soil nematode Caenorhabditis elegans exhibited higher mortality in 24 h in the presence of VOCs. These results suggest the broad-spectrum activity of Galactomyces fungi against various plant pathogens and the potential to use VOCs from Galactomyces as biocontrol agents.

• Journal of Ginseng Research
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• v.38 no.2
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• pp.136-145
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• 2014

Background: This study aimed to develop a biocontrol system for ginseng root rot caused by Fusarium cf. incarnatum. Methods: In total, 392 bacteria isolated from ginseng roots and various soils were screened for their antifungal activity against the fungal pathogen, and a bacterial isolate (B2-5) was selected as a promising candidate for the biocontrol because of the strong antagonistic activity of the bacterial cell suspension and culture filtrate against pathogen. Results: The bacterial isolate B2-5 displayed an enhanced inhibitory activity against the pathogen mycelial growth with a temperature increase to $25^{\circ}C$, produced no pectinase (related to root rotting) an no critical rot symptoms at low [$10^6$ colony-forming units (CFU)/mL] and high ($10^8CFU/mL$) inoculum concentrations. In pot experiments, pretreatment with the bacterial isolate in the presumed optimal time for disease control reduced disease severity significantly with a higher control efficacy at an inoculum concentration of $10^6CFU/mL$ than at $10^8CFU/mL$. The establishment and colonization ability of the bacterial isolates on the ginseng rhizosphere appeared to be higher when both the bacterial isolate and the pathogen were coinoculated than when the bacterial isolate was inoculated alone, suggesting its target-oriented biocontrol activity against the pathogen. Scanning electron microscopy showed that the pathogen hyphae were twisted and shriveled by the bacterial treatment, which may be a symptom of direct damage by antifungal substances. Conclusion: All of these results suggest that the bacterial isolate has good potential as a microbial agent for the biocontrol of the ginseng root rot caused by F. cf. incarnatum.

• 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.