• The Korean Journal of Soil Zoology
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• v.9 no.1_2
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• pp.1-5
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• 2004

Insect pests and their natural enemies of Hibiscus Linne (Malvaceae) were investigated from March 2002 to November 2004. Fourteen insect pest species of 9 families in 5 orders were collected from Hibiscus syriacus: 5 species in Homoptera, 3 species in Lepidoptera, 2 species in Coleoptera, 1 species in Orthoprera, 1species in Hemiptera, 1 spedies in Acarina, and 1 species in Stylommatophora. Especially, Aphis gossypii Glover (Aphididae), Anomis megogona Walker(Noctuidae) and Tetranychus urticae Koch (Tetranychidae) were very important species because of their increasing daminge. The highest donsities were observed from May to June in August in Tetranychus urticae. As the enemies and ento-mopathogens of insect pests on Hibiscus syriacus, 1 species of bacteria, 3 species of fungi, 1 species of fungi, 1 species of Hemiptera, 1 species of Coleoptera, 2 species of Hymenopetera, 2 species of Diptera, and 1 species of Acarina were investigated. As the predators and parasitoids of Aphis gossypii, Aphidoletes aphidoletes aphidimyza Rondani (Cecidomyiidae), Meliscaeva cinctella Zetterstedt (Syrphidae), Harmonia axyridis Pallas (Coccinellidae), and Aphidius gifuensis Ashmead (Braconidae), entomopathogenic fungi, Vericillium lecani naturalis strain (Moniliaceae) and Beauveria bassiana naturalis strain strain (Hypocreaceae) were observed and Bacillus thuringiensis naturalis strain (Bacillaceae), B. bassiana, Metarhizium anisopliae naturalis strain (Hypocreaceae), predators of Tetranychus urticae, Amblyseius sp. (Phytoseiidae), and Orius sp. (Anthocoridae) were observed.

• International Journal of Industrial Entomology and Biomaterials
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• v.26 no.2
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• pp.89-94
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• 2013

Entomopathogenic fungal species have been investigated for their potential use as biological control agents owing to their natural role as insect pathogens. These fungi produce a wide range of secondary metabolites with high therapeutic values, such as antibiotics and cytotoxic substances. To evaluate the antibacterial activity of entomopathogenic fungi, 10 isolates from Korean soil were selected and tested for their activity against Escherichia coli by using fungal culture filtrates. Antibacterial activity was assessed using a two-step process: (1) a screening assay for the selection of fungal isolates and (2) a quantitative assay to evaluate the activity of select fungi. Although 4 fungal isolates were selected through the screening assay, only 3 fungal isolates, from Beauveria bassiana and Metarhizium anisopliae, showed high antibacterial activity according to the quantitative assay. The antibacterial activity of selected fungal culture filtrates was stable when exposed to heat and proteolytic enzyme treatments, which indicated that the antibacterial compound is not a protein. These entomopathogenic fungal metabolites might be useful as a source for bacterial control and in the pharmaceutical industry.

• International Journal of Industrial Entomology and Biomaterials
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• v.30 no.2
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• pp.64-74
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• 2015

Beetles Protaetia brevitarsis seulensis Kolbe (Coleoptera: Cetoniidae) and Allomyrina dichotoma Linn. (Coleoptera: Scarabaeidae) are widely used in traditional medicine, and the number of insect-rearing farms is increasing in South Korea. The purpose of this study was to establish a multiplex PCR-based assay for rapid simultaneous detection of multiple pathogens causing insect diseases. Six insect parasites such as fungi Beauveria bassiana (Bals.-Criv.) Vuill. (Hypocreales: Cordycipitaceae) and Metarhizium anisopliae (Metschn.) Sorokin (Hypocreales: Clavicipitaceae), bacteria Bacillus thuringiensis Berliner (Bacillales: Bacillaceae), Pseudomonas aeruginosa Migula (Pseudomonadales: Pseudomonadaceae), and Serratia marcescens Bizio (Enterobacteriales: Enterobacteriaceae), and Oryctes rhinoceros nudivirus were chosen based on the severity and incidence rate of insect diseases in South Korea. Pathogen-specific primers were designed and successfully applied for simultaneous detection of multiple infectious agents in farm-bred insects P. b. seulensis and A. dichotoma using multiplex PCR and high resolution capillary electrophoresis. Our results indicate that multiplex PCR is an effective and time-saving method for simultaneous detection of multiple infections in insects, and the QIAxcel capillary electrophoresis system is useful for quantitative evaluation of the individual impact of each infectious agent on the severity of insect disease. The approach designed in this study can be utilized for rapid and accurate diagnostics of infection in insect farms.

• International Journal of Industrial Entomology and Biomaterials
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• v.33 no.1
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• pp.1-5
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• 2016

Beauveria bassiana is a common fungal pathogen of Protaetia brevitarsis larvae, and although it is less common than Metarhizium anisopliae , the pathogen still poses a great risk to humans and animals that consume infected insects, owing to B. bassiana's production of toxins like beauvericin and mycotoxin. Interestingly, the beneficial microorganism Saccharomyces cerevisiae possesses antifungal properties. In the present study, we found that S. cerevisiae inhibited the growth of B. bassiana by 97% and that S. cerevisiae failed to harm P. brevitarsis when administered via intracoelomic injection (1×10⁷ cfu/mL). In addition, we also found that S. cerevisiae consumption increased the survival time of percutaneously infected P. brevitarsis larvae by 5 d and reduced the mortality of infected larvae by 12%. Therefore, S. cerevisiae is expected to be useful in the prevention and control of B. bassiana in the production of P. brevitarsis larvae.

• Mycobiology
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• v.45 no.3
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• pp.192-198
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• 2017

The green peach aphid (Myzus persicae), a plant pest, and gray mold disease, caused by Botrytis cinerea, affect vegetables and fruit crops all over the world. To control this aphid and mold, farmers typically rely on the use of chemical insecticides or fungicides. However, intensive use of these chemicals over many years has led to the development of resistance. To overcome this problem, there is a need to develop alternative control methods to suppress populations of this plant pest and pathogen. Recently, potential roles have been demonstrated for entomopathogenic fungi in endophytism, phytopathogen antagonism, plant growth promotion, and rhizosphere colonization. Here, the antifungal activities of selected fungi with high virulence against green peach aphids were tested to explore their potential for the dual control of B. cinerea and M. persicae. Antifungal activities against B. cinerea were evaluated by dual culture assays using both aerial conidia and cultural filtrates of entomopathogenic fungi. Two fungal isolates, Beauveria bassiana SD15 and Metarhizium anisopliae SD3, were identified as having both virulence against aphids and antifungal activity. The virulence of these isolates against aphids was further tested using cultural filtrates, blastospores, and aerial conidia. The most virulence was observed in the simultaneous treatment with blastospores and cultural filtrate. These results suggest that the two fungal isolates selected in this study could be used effectively for the dual control of green peach aphids and gray mold for crop protection.

• Journal of Microbiology
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• v.44 no.6
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• pp.622-631
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• 2006

In this study we purified a fibrinolytic enzyme from Cordyceps militaris using a combination of ion-exchange chromatography on a DEAE Sephadex A-50 column, gel filtration chromatography on a Sephadex G-75 column, and FPLC on a HiLoad 16/60 Superdex 75 column. This purification protocol resulted in a 191.8-fold purification of the enzyme and a final yield of 12.9 %. The molecular mass of the purified enzyme was estimated to be 52 kDa by SDS-PAGE, fibrin-zymography, and gel filtration chromatography. The first 19 amino acid residues of the N-terminal sequence were ALTTQSNV THGLATISLRQ, which is similar to the subtilisin-like serine protease PR1J from Metarhizium anisopliae var. anisopliase. This enzyme is a neutral protease with an optimal reaction pH and temperature of 7.4 and $37^{\circ}C$, respectively. Results for the fibrinolysis pattern showed that the enzyme rapidly hydrolyzed the fibrin $\alpha$-chain followed by the $\gamma$-$\gamma$ chains. It also hydrolyzed the $\beta$-chain, but more slowly. The A$\alpha$, B$\beta$, and $\gamma$ chains of fibrinogen were also cleaved very rapidly. We found that enzyme activity was inhibited by $Cu^{2+}$ and $Co^{2+}$, but enhanced by the additions of $Ca^{2+}$ and $Mg^{2+}$ ions. Furthermore, fibrinolytic enzyme activity was potently inhibited by PMSF and APMSF. This enzyme exhibited a high specificity for the chymotrypsin substrate S-2586 indicating it's a chymotrypsin-like serine protease. The data we present suggest that the fibrinolytic enzyme derived from the edible and medicinal mushroom Cordyceps militaris has fibrin binding activity, which allows for the local activation of the fibrin degradation pathway.

• The Korean Journal of Mycology
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• v.25 no.4 s.83
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• pp.239-252
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• 1997

Thirty three entomopathogenic species belonging to twelve genus were collected throughout 16 collecting sites from 1990 to 1996. Among those collected species, 14 unrecorded species such as Cordyceps bifusispora, C. martialis, C. oxycephala, C. paludosa, C. pentatomi, C. rosea, C. ryogamiensis, Shimizuomyces paradoxa, Akanthomyces aculeatus, Polycephalomyces ramosus, Tilachlidiopsis nigra were added to Korean entomopathogenic species through this study. In nature, occurrence of Cordyceps nutans, C. sphecocephala and Paecilomyces tenuipes were from early June to late September. On the other hand, C. militaris, C. kyushuensis and C. pruinosa were mainly found from mid July to mid August when relative humidity are increased. Nine species of the genus Cordyceps including C. bifusispora and four deuteromycetous species were isolated. As a result of cultural test using six Cordyceps species, anamorph of C. militaris, C. kyushuensis were proved as Verticillium sp. C. pruinosa as Acremonium sp., C. sphecocephala as Hymenostilbe sp. and C. scarabaeicola as Beauveria sp., respectively.