• Journal of Forest and Environmental Science
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• v.34 no.5
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• pp.376-381
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• 2018

For the control of oak wilt caused by Raffaelea quercus-mongolicae, an antifungal microorganism, Streptomyces blastmyceticus, was used as a potential agent. Culture suspension of S. blastmyceticus was injected into Quercus mongolicae in the research forest of Kangwon National University by $ChemJet^{(R)}$ trunk injection and Macro-infusion at root flare injection. $Alamo^{(R)}$ (a.i., propiconazole 14.5%), a fungicide currently used for the control of oak wilt in USA, was also treated by both methods to compare the efficacy. For preventive efficacy, culture suspension of the pathogen was inoculated at 1 month after injection of either agent. Tested trees were cut at 3 months after treatment, stained with 1% Fuchsin acid, and then non-conductive area (NCA) and re-isolation frequency (RIF) of oak wilt fungus were compared among treatments. While NCA was the highest as 47.3% in pathogen only treatment, it was the lowest as 16.0% in sterilized water treatment by Macro-infusion. NCAs of Alamo treatment by Macro-infusion and ChemJet injection were 25.3% and 32.1%, respectively. NCA of S.blastmyceticus treatment by ChemJet injection was 32.3%, similar with Alamo treatment's by ChemJet injection. All treatments by either injection method showed significantly lower NCA compared to the pathogen only treatment. These results indicate that S. blastmyceticus injection shows the preventive efficacy against oak wilt disease by suppressing the growth of pathogen injected. NCA of Macro-infusion injection of sterilized water was lower as 16.0%, compared to 21.3% of ChemJet injection. It means that Macro-infusion is more effective in translocation of sterilized water than ChemJet injection by even distribution. RIF from wood discs of treated trees showed high in pathogen only treatment, but relatively low in S. blastmyceticus treatment. RIF results were correlated with NCA results. From the above results, it was confirmed that S. blastmyceticus showed preventive efficacy against oak wilt disease by ChemJet trunk or Macro-infusion at root flare injection.

• Journal of Forest and Environmental Science
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• v.34 no.5
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• pp.395-404
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• 2018

Since the mass mortality of Quercus mongolica has been first reported in Gyeonggi province at 2004, the disease spread rapidly over Korean peninsula annually. Ambrosia beetle (Platypus koryoensis) was known as the insect vector of oak wilt fungus, Raffaelea quercus-mongolicae, and control methods of the disease had mainly been focused on eradication of insect vector. However, for the efficient management of the disease, combined control methods for both of the pathogenic fungus and insect vector are strongly required. As one of the efforts to suppress the pathogenic fungus, antifungal activities of Streptomyces isolated from oak forest soil were assayed in this study. Optimum culture condition for the selected isolates was also studied, As a result, Streptomyces blastmyceticus cultured in PDB (Potato Dextrose Broth) at $25^{\circ}C$ for 1 week showed the strongest antifungal activity against oak wilt fungus. Mycelial growth inhibition rates (MGIRs) of Streptomyces isolates were compared on culture media supplemented with heated and unheated culture filtrates of S. blastmyceticus. MGIRs on culture media with unheated culture filtrates were generally higher than those on culture media with heated culture filtrates. Antagonistic mechanism to get involved in the inhibition of hyphal growth and spore formation of the pathogen is due to the antifungal metabolites produced by Streptomyces. This study will provide the fundamental information in developing biocontrol agents for the environment-friendly management of oak wilt disease.

• Mycobiology
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• v.47 no.3
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• pp.329-334
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• 2019

Streptomyces blastmyceticus strain 12-6 was isolated from a forest soil sample of Cheonan area on the basis of strong antifungal activities against plant pathogenic fungi. Butanol extracts of the cultural filtrates were active against C. acutatum, C. coccodes, C. gloeosporioides, F. oxysporum, and T. roseum. Active fractions were prepared by thin layer chromatography using silica gel plate; 12-6-2 ($R_f$ 0.36), 12-6-3 ($R_f$ 0.44). Scanning electron microscopy showed that the active fractions caused a change in surface texture of fungal spores from smooth surface to wrinkled surface. The lethal effect on the spores of the active fractions varied from 56% to 100%. It was shown that the spores of C. acutatum were more sensitive to the antifungal fractions than the spores of F. oxysporum. Fluorescence staining using TOTO-1 indicated that the antifungal fractions could make the spores more sensitive to the fluorescence dye. Thus, it was suggested that antifungal agents prepared in this study exhibited the antifungal activity by damaging the plasma membrane of both fungal spores and hyphae. Identification of antifungal agents in the active fraction using GC-MS analysis revealed the presence of cyclo-(Leu-Pro) and 9-octadecenamide as major components that have already been known as antifungal substances.