• Mycobiology
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• v.48 no.6
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• pp.484-494
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• 2020

Oak wilt disease caused by Raffaelea quercus-mongolicae has emerged obviously in Korea. We selected antifungal isolates against R. quercus-mongolicae among 368 endophytic fungal isolates from different parts of oak and pine trees. The experiment was conducted in the primary and secondary screenings by dual culture test. The antifungal activity of the selected isolates was assessed in culture filtrate test based on the inhibition rates in mycelial growth, sporulation, and spore germination of oak wilt fungus. Five isolates, E089, E199, E282, E409 and E415, showed strong antifungal activity in culture filtrate test, and their antifungal activity decreased on the culture media supplemented with heated culture filtrate. Higher mycelial growth inhibitions on the unheated media were recorded in E409 (Colletotrichum acutatum), E089 (Daldinia childiae), E415 (Alternaria alternata) and E199 (Daldinia childiae) with the inhibition rates of 79.0%, 70.1%, 68.9% and 64.5%, respectively. These isolates also had the higher sporulation inhibitions on unheated media with the rates of 96.8%, 84.2%, 82.8% and 80.5%, respectively. The spore germination of the oak wilt fungus was completely inhibited by E282 (Nectria balsamea) on both unheated and heated media. These results showed that a higher number of potent antifungal isolates against oak wilt fungus was isolated from the petiole compared to the other parts. This study could contribute to the development of biological control approaches for the management of oak wilt disease caused by R. quercus-mongolicae.

• 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.

• 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.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.18-18
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• 2015

Wilt disease in Oak trees occurs during summer season in Korea. Mass attack of trees by an ambrosia beetle (Platypus koryoensis) was the characteristic feature before appearance of the wilting symptoms. Raffaelea sp. caused the discoloration of xylem area called as wound heartwood. Raffaelea sp. was observed both on the body surfaces and inside the mycangia of the beetle Platypus sp. The scanning electron microscope (SEM) analysis showed that fungal spores were present within the wall of gallery and vessels that formed tyloses. The results revealed that the water movement in vessels was blocked as the fungus started to grow which caused the formation of tyloses thereby resulting wilt symptoms. We found that both female and male beetle Platypus sp. had fungi on their bodies and their large and small mycangia. This study confirmed that the fungus was transferred to oak trees by Platypus sp.

• Journal of Forest and Environmental Science
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• v.33 no.4
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• pp.342-347
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• 2017

Since the first report of the oak wilt disease at 2004 in Korea, the disease distributed over Korean peninsula and are still giving severe damages. The management of oak wilt disease in Korea has mainly focused on the control of insect vector, Platypus koryoensis. Neverthless the effective method for evaluating the pathogenicity of the pathogen, Raffaelea quercus-mongolicae (Rqm), and for screening chemical or biological agents with strong inhibitory activity against the pathogen, is absolutely necessary, an reliable method is not available so far. This study was conducted to develop the effective method for evaluating the pathogenicity of Rqm in oak trees. The culture suspensions of Rqm were artificially injected to the saplings of Quercus acutissima by using ChemJet tree injector. Three months after treatments, the treated saplings were cut and dipped into 1% fuchsin acid solution. There were significant differences in non-conductive area (%), discoloration area (%) and vertical discoloration length between the pathogen-injected and distilled water-injected control treatments. These results indicated that the pathogen is the causal agent for the dysfunction of water conductive tissue, which will finally result in wilt symptom. Re-isolation of the pathogen and PCR detection using specific primers for the pathogen also confirmed the presence of Rqm in the sapwood chips of the pathogen-injected saplings. These observations would be greatly applied to other related researches for evaluating the pathogenicity of tree wilt pathogens and biocontrol efficacy of the selected antagonistic microorganisms, in case that the wilt symptom is not easily shown by artificial inoculation of the causal agent.

• Journal of Microbiology and Biotechnology
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• v.19 no.8
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• pp.760-764
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• 2009

In this study, we investigated the antifungal activity of three different forms of silver nanoparticles against the unidentified ambrosia fungus Raffaelea sp., which has been responsible for the mortality of a large number of oak trees in Korea. Growth of fungi in the presence of silver nanoparticles was significantly inhibited in a dose-dependent manner. We also assessed the effectiveness of combining the different forms of nanoparticles. Microscopic observation revealed that silver nanoparticles caused detrimental effects not only on fungal hyphae but also on conidial germination.

• International Journal of Industrial Entomology and Biomaterials
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• v.18 no.2
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• pp.125-129
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• 2009

Entomopathogenic fungi were isolated directly from a cadaver of adult Monochamus alternatus supporting fungal sporulation, using a semi-selective medium and then screened several fungal colonies. The pathogenicity of each fungus was tested using oak longicorn beetle, Moechotypa diphysis, as substitutive insect. As the result, only one of them showed high pathogenicity against M. diphysis, with up to 100% mortality within 21 days of inoculation. Selected fungus was named as MaW1 and identified by Beauveria bassiana using microscopic examination and DNA analysis. Pathogenicity was also evaluated to M. alternatus.