• The Korean Journal of Mycology
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• v.21 no.2
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• pp.94-105
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• 1993

The genus Cordyceps known as an insect parasite forms a sclerotium in insect bodies and then produces perithecia on the single or multiple stromata produced from sclerotium. Collected Cordyceps were identified into 5 species: Cordyceps militaris, C. nutans, Cordyceps sphecocephala, Isaria japonica, and Torrubiella sp. The fruit bodies of Cordyceps in petri-dish cover were fixed by tape and put the lid on water agar plates to isolate these collected Cordyceps. The germinated spores were transferred from water agar to Potato dextrose agar(PDA) after six hours. Mycelial growth of C. nutans and C. militaris was the most successful on Hamada media and was also good on Complete media and PDA. Mannose as a carbon source was good for two species and Glutamic acid as a nitrogen source was satisfactory to C. militaris and Asparagine gave a good result to C. nutans. C. militaris and C. nutans showed similar mycelial growth rate on the media that contained thiamine-HCI, biotine or nicotinic acid as a vitamine. When conidia of C. nutans were inoculated to insects, mortality was high in Artogeia napi L, Hemiptera, Plutella xylostella and 50% in Orthoptera, 12% in Acantholyda posticalise M, but not Agelastica coerulea B. in Aphididae, C. nutans was collected from only Hemiptera in nature, but killing effect on other insects was proved. Mycelial growth and fruit-body formation were good on the media that consist of rice powder 5g, wheat flour 5g, water 100ml, but formed fruit-body was not complete stromata but a mass of conidia according to results of observing microscope.

• Korean journal of applied entomology
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• v.53 no.3
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• pp.271-280
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• 2014

Some bacterial metabolites of Xenorhabdus nematophila (Xn) inhibit phospholipase $A_2$ ($PLA_2$) activity to shutdown eicosanoid biosynthesis in target insects. However, little has been known about the target insect $PLA_2$ of these bacterial metabolites. Eight bacterial metabolites identified in Xn culture broth exhibited significant insecticidal activities against larvae of both lepidopteran species of Plutella xylostella and Spodoptera exigua. Moreover, these bacterial metabolites significantly enhanced insecticidal activities of Bacillus thuringiensis (Bt). To determine target $PLA_2$, we cloned and over-expressed cellular $PLA_2$ ($SecPLA_2$) of S. exigua. Purified $SecPLA_2$ catalyzed phospholipids derived from the fat body and released several polyunsaturated fatty acids. Most Xn metabolites significantly inhibited $SecPLA_2$ activity, but were different in their inhibitory activities. There was a positive correlation between the inhibition of $SecPLA_2$ and the enhancement of Bt insecticidal activity. These results indicate that $SecPLA_2$ is a molecular target inhibited by Xn metabolite.

• Korean Journal of Microbiology
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• v.48 no.4
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• pp.284-292
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• 2012

To date, chemical control remains the most common way to reduce beet armyworm (Spodoptera exigua) populations. However, this insect has become more tolerant or resistant to many chemical insecticides and the insect larvae usually hide inside hollow, tube-like leaves of host plant so they were difficult to kill by spraying insecticides. The use of viral and bacterial insecticide to solve these problems has not been successful because of their novel feeding habit. To overcome these problems, in this study, the biological characteristics and virulence of an entomopathogenic fungus isolated from the cadaver of larvae beet armyworm were investigated. Isolated entomopathogenic fungus was identified as Nomeraea rileyi (Farlow) Samson by morphological examinations and genetic identification using sequences of the ITS, ${\beta}$-tubulin gene and EF1-${\alpha}$ regions. This fungus was named as N. rileyi SDSe. Virulence tests against 3rd larvae of beet armyworm were conducted with various conidial suspensions from $1{\times}10^4$ to $10^8$ conidia/ml of N. rileyi SDSe in laboratory conditions. Mortality rate of beet armyworm showed from 20 to 54% and the virulence increased with increasing conidial concentrations. Although N. rileyi SDSe showed low mortality rate against beet armyworm, it is expected that N. rileyi SDSe will be used effectively in the integrated pest management programs against the beet armyworm.

• Microbiology and Biotechnology Letters
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• v.16 no.5
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• pp.389-392
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• 1988

A clone pSL 2-1, which is a recombinant plasmid believed to contain the mosquitocidal crystal-line protein gene of the Bacillus sphaericus 1593, was expressed in Escherichia coli JM83 and the product of the clone was purified and identified. The unsolubilized mosquitocidal crystal proteins from the B. sphaericus had formed 43, 58, 64, 100, 113, and 130 Kd bands in the SDS-polyacrylamide gel, but the NaOH-solublized proteins at pH 12 formed 2 protein bands of 43- and 64Kd in the gel because the larger protein (precursor) bands were cleaved. The products of the pSL 2-1 clone was purified by Sephadex G-200 and only the fractions having lethal activity to the 3rd in-star larvae of mosquito Culex pipiens were analyzed by the gel. The only single protein band of 42 Kd toxic to the larvae was formed. The major toxic protein being produced from the B. sphaericus 1593 and the pSL 2-1 clone was found to be the 42 Kd.

• Korean journal of applied entomology
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• v.55 no.4
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• pp.319-327
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• 2016

Chemical pesticides have been used to control persimmon pests, however the overuse of the pesticides caused insect resistance, followed by failure in pest management and residual problems. Herein we investigate the potential of eco-friendly organic pesticides (EFOP) on the control persimmon pests, Stathmopoda masinissa (persimmon fruit moth) and Riptortus pedestris (bean bug). Ten commercially available plant-derived organic pesticides and one microbial pesticide were sprayed on the target insects in laboratory conditions. The chemical pesticide, buprofezin+dinotefuran wettable powder served as a positive control. In the first bioassay against persimmon fruit moth, alternatively Plutella xylostella larvae were used due to the lack of persimmon fruit moth population from fields, and three organic pesticides showed high control efficacy, such as pyroligneous liquor (EFOP-1), the mixture of Chinese scholar tree extract, goosefoot and subtripinnata extracts (EFOP-2) and Bacillus thuringiensis subsp. aizawai NT0423 (EFOP-11). When the three selected organic pesticides were treated on the persimmon fruit moths, the EFOP-2 treatment showed the highest control efficacy: 27.7% (5 days), 13.3% (7 days) and 6.7% (10 days) of survival rates. In the bioassay against bean bugs, the mixture of Chinese scholar tree, goosefoot and subtripinnata extracts (EFOP-2 and EFOP-9) and the extracts of sophora and derris (EFOP-10) showed high control efficacy, particularly the highest in the treatment of EFOP-2: 20.0% (5 days) and 16.7% (10 days) of survival rates. These results suggest that the mixture of Chinese scholar tree, goosefoot and subtripinnata extracts (EFOP-2) has high and multiple potential in the management of the persimmon pests.

• Korean journal of applied entomology
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• v.59 no.1
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• pp.41-54
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• 2020

Two mosquito species were collected in still-water near farming area in Andong, Korea. Based on morphological characters, these two mosquitoes were identified as Aedes koreicus and Culex vagans, respectively. DNA barcode analyses supported the identification. An entomopathogenic bacterium, Bacillus thuringiensis subsp. israelensis (BtI), exhibited insecticidal activities against the two mosquito species and its virulence was more potent than that of B. thuringiensis subsp. kurstaki. It has been known that the bacterial metabolites of Xenorhabdus spp. suppress insect immunity and enhance pathogenicity of B. thuringiensis. This study tested the effect of the bacterial culture broth of Xenorhabdus spp. on enhancing BtI pathogenicity. Among three Xenorhabdus spp., culture broth of X. ehlersii (Xe) was relatively effective to enhance BtI pathogenicity against both mosquito species. Indeed, organic extracts of Xe culture broth suppressed the hemocyte-spreading behavior, suggesting the presence of immunosuppressant in the culture broth. These results suggest a formulation of BtPlus by mixing BtI spore and Xe culture broth to be applied to control the two mosquito species.

• The Korean Journal of Ecology
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• v.22 no.5
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• pp.255-263
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• 1999

Entomopathogenic nematodes were isolated through the investigation of soil samples from various biotopes in south Korea, the efficiency of isolation for highly pathogenic nematodes to silkworms (Bombyx mori) was as high as 28 %. Twenty-eight strains of nematodes, selected among 100 samples by silkworms were confirmed the pathogenicity, multiplicity, and tolerance against various condition of preservation. Pathogenicity of the nematode isolates to agricultural and environmental pests such as Calliphora vomitoria, Pseufaletia separata, Palomena angulosa, and Melolontha incana were high. Mortality was varied from 20 to 100% by the pest insects and nematode strains. The high detectablity of entomopathogenic nematodes resulted from the methods of collection for soil samples within 10 cm depth after eliminating dried soil surface and the use of silkworm trap. High population of entomopathogenic nematodes represented the strong activity and broad action radius in the environment. Most of the nematode isolates were successfully cultured on the silkworm host as well as on artificial media, and proved their potential for the use of biological control agent.

• Korean journal of applied entomology
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• v.55 no.3
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• pp.257-266
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• 2016

Subtropical insect pests expand their habitats by migration to temperate zones along with global climate change. A subtropical insect pest, Maruca vitrata, is infesting leguminous crops including azuka beans in Korea and gives significant economic damages. Its great genetic variation raised an issue of the origin of a Korean M. vitrata population. To understand the genetic character of the Korean population, its cytochrome oxidase subunit 1 (cox 1) gene was sequenced and phylogenetically analyzed with other regional populations. The world populations of M. vitrata were grouped into three clusters: Asia-African, American, and Oceanian. The Korean population was classified into Asia-African cluster. To characterize the insecticide susceptibility of the Korean population, seven different insecticides (4 neutoxic insecticides, 1 insect growth regulator, 2 biopesticides) were assessed. Young larvae of M. vitrata were relatively susceptible to all tested insecticides. However, old larvae were much less susceptible than young larvae. No test insecticides effectively (> 50%) killed the old larvae of M. vitrata within 7 days.

• The Korean Journal of Pesticide Science
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• v.12 no.2
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• pp.184-191
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• 2008

Entomopathogenic bacteria (Xenorhabdus nematophila, X. sp. and Photorhabdus temperata subsp. temperata) isolated from entomopathogenic nematodes express potent insecticidal activity in insect hemocoel. They are also known to suppress insect immune mediation by inhibiting phospholipase $A_2$, leading to host immunosuppression. This study analyzed effects of their cultured broths on inhibiting insect immunosuppression. For this, we removed all bacterial cells using $0.2\;{\mu}m$ pore sized membrane from the bacteria-cultured broth. All three sterilized cultured media, in dose-dependent manners, significantly inhibited hemocyte-spreading behavior of 5th instar larvae of Spodoptera exigua. However, they showed differential inhibitory activities among different bacterial species, in which X. nematophila showed the most potent inhibitory activity. This immunosuppressive effect was applied to increase the pathogenicity of Bacillus thuringiensis (Bt). All three bacterial cultured broths including bacterial cells significantly potentiated Bt pathogenicity against young S. exigua larvae when each of them was orally administered in a mixture of low dose of Bt. Finally, we tested the effect of oral administration of the cultured media containing the immunosuppressive compound(s) secreted by the bacteria. The membrane-sterilized cultured broths were mixed with the low dose of Bt and then orally administered to the young S. exigua. Only the cultured medium of X. nematophila showed increase of Bt pathogenicity. These results indicated that the; cultured media of the three bacteria possessed immunosuppressive factor(s), which may act to potentiate Bt toxicity to young S. exigua larvae.

• Korean journal of applied entomology
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• v.56 no.1
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• pp.19-28
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• 2017

Polydnaviruses (PDVs) are a group of double-stranded DNA viruses symbiotic to some endoparasitoid wasps. Cotesia plutellae bracovirus (CpBV) is a PDV symbiotic to an endoparasitoid wasp, C. plutellae, parasitizing young larvae of Plutella xylostella. An early expressed gene, CpBV-ELP1, plays an important role in the parasitism by suppressing host cellular immunity by its cytotoxic activity against hemocytes. This study aimed to test its oral toxicity against insect pest by expressing it in a recombinant tobacco plant. A recombinant CpBV-ELP1 protein was produced using a baculovirus expression system and secreted to cell culture medium. The cell cultured media were used to purify CpBV-ELP1 by a sequential array of purification steps: ammonium sulfate fractionation, size exclusion chromatography, and ion exchange chromatography. Purified rCpBV-ELP1 exhibited a significant cytotoxicity against Spodoptera exigua hemocytes. CpBV-ELP1 was highly toxic to the fifth instar larvae of S. exigua by injection to hemocoel. It also showed a significant oral toxicity to fifth instar larvae of S. exigua by a leaf-dipping assay. CpBV-ELP1 was cloned into pBI121 vector under CaMV 35S promoter with opaline synthase terminator. Resulting recombinant vector (pBI121-ELP1) was used to transform Agrobacterium tumefaciens LBA4404. The recombinant bacteria were then used to induce callus of a tobacco (Nicotiana tabacum Xanthi) leaves and subsequent generation (T1) plants were selected. T1 generation tobacco plants expressing CpBV-ELP1 gave significant insecticidal activities against S. exigua larvae. These results suggest that CpBV-ELP1 gene can be used to control insect pests by constructing transgenic crops.