• Research in Plant Disease
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• v.20 no.4
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• pp.270-274
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• 2014

Division of Forest Diseases & Insect Pests, Korea Forest Research Institute, Seoul 130-712, Korea The objective of this study was to evaluate the virulence of geographically different isolates of oak wilt pathogen, Raffaelea quercus-mongolicae and other Raffaelea species. In this study, mature trees of Quercus mongolica were inoculated with the various isolates of Raffaelea spp. and their virulence was evaluated by measuring the extent of sapwood discoloration resulting from the inoculation. The average length of discolored sapwood in a lateral direction was longest in the trees inoculated with the isolates from Korea (8.69 cm) followed by R. quercivora (7.51 cm) and the other Raffaelea spp. (3.35 cm). The lateral length of discolored sapwood caused by the inoculation with Korean strains varied from 4.71 to 14.90 cm indicating their differences in virulence. The area of discolored sapwood caused by the inoculation with Raffaelea spp. varied from 1.57 to $8.42cm^2$ indicating their differences in virulence. Based on the length and area of the discolored sapwoods, isolated YY and wj43 appeared to have the highest virulence among all the Raffaelea isolates tested. Each of the two isolates was obtained from Gangwon Province and Jeonbuk Province, respectively.

• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.66-69
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• 2012

Five strains out of 200 candidate strains (SG 1-9, 1-12, SG 2-8, 2-10, and 2-17) were selected to determine their antifungal activity against Raffaelea quercus-mongolicae. The 16S rDNA sequences of the five strains were determined by sequencing analysis and analyzed by the homology of the blast program at NCBI. The homology search showed that SG 1-9 and 1-12 had a 98% homology with Streptomyces cinnamoneus and 98% homology with Burkholderia cepacia, while SG 2-8, 2-10, and 2-17 had a 99% homology with Streptomyces fradiae, a 97% homology with Staphylococcus epidermidis, and a 99% homology with Staphylococcus epidermidis. Out of the five selected strains, organic extract and protein extracts of SG2-17 strain broth were employed to determine antifungal activity against Raffaelea quercus-mongolicae. The organic extract exhibited antifungal activity, but the protein extracts did not demonstrate such an activity. Three organic solvents, butanol, benzene, and ethyl acetate, were also used for determination of antifungal activities. The activity measurements revealed that benzene extract possessed the greatest inhibitory effect on the growth of Raffaelea quercus-mongolicae, with the next highest being butanol extract, and ethyl acetate extract being the lowest.

• Research in Plant Disease
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• v.16 no.3
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• pp.219-223
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• 2010

Recently, the oak wilt diseases especially on Quercus mongolica, have been increasing in various districts of Korea. A collection of 38 strains of the oak wilt pathogen Raffaelea qeurcus-mongolicae and R. quecivora isolated from Quercus spp. in Korea and Japan was characterized by $\beta$-tubulin gene sequence and randomly amplified polymorphic DNA (RAPD) analysis. In cluster analysis based on $\beta$-tubulin gene sequence the strains were divided into 4 clusters, of which clusters 2 and 4 were composed of Japanese strains except for one Korean strain. RAPD analysis showed that they were also effectively differentiated by a strong RAPD fragments. On the basis of the two genetic analysis, significant differences were detected between Korean strains and Japanese strains.

• Research in Plant Disease
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• v.17 no.2
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• pp.136-141
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• 2011

In other previous in vitro tests, the hyphal growth of Raffaelea quercivorus was inhibited by the treatments of various concentration of nano-silver. In this field tests, treatment of different concentrations of nano-silver to oak trees in Cheolwon and Hongcheon sites showed the inhibition effects against wilt disease caused by Raffaelea quercivorus. However, nano-silver-treated oak tree in Chuncheon site showed mild wilt symptoms with no phytotoxicity. Scanning electron microscope (SEM) observation confirmed that the spore and hypha of Raffaelea sp. inside the vessels were damaged by nano-silver. This result indicates the application of nanosilver is effective for control of Raffaelea quercivorus in the field.

• 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.

• Mycobiology
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• v.42 no.2
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• pp.210-214
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• 2014

In Korea, mass mortality of Quercus mongolica trees has become obvious since 2004. Raffaelea quercus-mongolicae is believed to be a causal fungus contributing the mortality. To evaluate the pathogenicity of the fungus to the trees, the fungus was multiple- and single-inoculated to the seedlings and twigs of the mature trees, respectively. In both the inoculations, the fungus was reisolated from more than 50% of inoculated twigs and seedlings. In the single inoculations, proportions of the transverse area of non-conductive sapwood at inoculation points and vertical lengths of discoloration expanded from the points were significantly different between the inoculation treatment and the control. In the multiple inoculations, no mortality was confirmed among the seedlings examined. These results showed that R. quercus-mongolicae can colonize sapwood, contribute to sapwood discoloration and disrupt sap flows around inoculation sites of Q. mongolica, although the pathogenicity of the fungus was not proven.

• 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.

• Korean journal of applied entomology
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• v.45 no.2 s.143
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• pp.113-117
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• 2006

Occurrence of the oak wilt disease (Raffaelea sp.) was confirmed in 2004 for the first time in Korea. By using pictorial keys, morphological characteristics of Platypus koryoensis (Murayama), a vector for this disease in Korea, was compared to Platypus quercivorus (Murayama), a vector of oak wilt in Japan. Additionally, suggestions were made for the future research of the vector of oak wilt disease in the northeastern Asia.

• Research in Plant Disease
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• v.20 no.4
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• pp.295-298
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• 2014

Oak wilt caused by Raffaelea quercus-mongolicae was first noticed in South Korea in 2004 and, ever since, its distribution and damage have been increasing. To screen a fungicide effective for oak wilt control by tree injection, laboratory and field experiments were conducted. Ten fungicides and one antibiotic were examined in vivo for their effectiveness in restricting the growth of R. quercus-mongolicae and R. quercivora (Japanese oak wilt pathogen) isolates. To the Korean isolates of R. quercus-mongolicae, chlorothalonil showed the highest fungicidal effects, followed by benomyl and propiconazole. To the Japanese one, propiconazole was highest in the fungicidal effectiveness, followed by benomyl and bitertanol. Propiconazole was selected for field-testing of its control efficacy because it showed good fungicidal effects in vitro and systemic activity. The control efficacy in the field was 87.5% in the first year of injection and 66.7% in the second year, indicating the fungicidal effects last at least over one year.

• 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.