• KSBB Journal
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• v.17 no.3
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• pp.276-282
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• 2002

Symbiotic bacteria with highly effective insecticidal activities were isolated and compared with their physiological characteristics from seven species of entomopathogenic nematodes belong to Steinernamatidae and Heterorhabditidae sp., and three of them were identified as Xenorhabdus nematophilus. Culture characteristics, insecticidal activities, pretense activities and fatty acid contents of various symbiotic bactierial isolates were also examined. In the case of cell growth and insecticidal activity, XR-PC and XR-MK were superior to other species when cultured in vitro. The insecticidal activity were highest at the early exponential growth phase, and gradually decreased with time. The protease activity of XR-DR was remarkable compared to other species. In the case of HE-HY, however the pretense activity increased in parallel with cell growth. Interestingly, the fatty acid patterns of Xenorhabdus nematophilus isolated from different emtomopathogenic nematode, showed remarkable differences in their contents of 12:0, 14:0, 16:1 cia 5 and 17:0 cyclo and hydroxy and branch factty acids were varied from 2% to 15% among total fatty acid contents.

• Korean journal of applied entomology
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• v.38 no.3
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• pp.255-260
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• 1999

Pathogenicity and multiplication of entomopathogenic nematode, Steinernema carpocapsae Weiser, ere analyzed in two insect hosts, Spodoptera exiga (Hubner) and Sp. litura(Fabricius). The estimated ${LC}^{50}s$(lethal concentration of the infective juveniles to kill 50% of the host insect population) were not different between tow insect species or among their developmental stages on the filter paper assay, thought the actual numbers of the infected nematodes were varied among them. The significant variation, however occurred in nematode multiplication between tow inset hosts. Temperature also gave sosignificatn effect on nematode multiplication rate that it took 6 days after infection at $25^{\circ}C$, but did 12 days at $20^{\circ}C$ to show the maximal nematode population peak($\approx$500,000 infective juveniles(IJ) in a 5th instar larva of Sp. litura and $\approx$-100,000 IJ in a 5th instar larva of Sp. exigua).

• Korean journal of applied entomology
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• v.45 no.2 s.143
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• pp.201-209
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• 2006

Five strains of Korean entomopathogenic nematodes (EPN), steinernematids and heterorhabditids were evaluated and tried in laboratory, pot, and vegetable greenhouses for environmentally friendly control of diamondback moth (DBM), Plutella xyiostella, from 2002 to 2005. LC$_{50}$ values of five EPN strains against DBM were different depending on nematode strain and DBM instar. LC$_{50}$ value of Steinernema carpocapsae GSN1 (GSN1) was the lowest representing 2.6$\sim$3.9 infective juveniles (Ijs, 3rd stage) to 2nd to 4th instars of DBM. Pathogenicity of five effective strains against DBM was different depending on nematode strain, concentration, application times, and vegetable in pot. The most effective nematode was GSN1. Steinernema spp. was more effective than Heterorhabditis spp. against DBM. Two or three times of applications of EPN were effective regardless of nematode strain and concentration in pot. Efficacy of EPN was different depending on vegetable species. Efficacy was higher on Chinese cabbage, red mustard, and Ssamchoo than that on cabbage, kale, and leaf broccoli. Efficacy of GSN1, Steinernema GSNUS-10, Steinernema CSNUS-14, and Heterorhabditis GSNUH-1 was variable depending on nematode strain, concentration, application times, and vegetable in greenhouse experiments. GSN1 was the most effective and 100,000 infective juveniles per m$^2$(=$1\times10^9$ Ijs/ha) resulted in higher efficacy. Three times of application of nematodes led to higher control efficacy than one or two applications. Efficacy of nematodes was higher on Chinese cabbage than cabbage or kale in greenhouse.

• The Korean Journal of Pesticide Science
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• v.20 no.4
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• pp.361-368
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• 2016

Japanese pine sawyer (JPS), Monochamus alternatus is a vector insect of pine wood nematode, Bersaphlenchus xylophilus in Korea and Japanese pine forest. This study was to evaluate the possibility of biological control of JPS with Korean entomopathogenic nematodes (EPNs). Korean EPNs (Heterorhabditis sp. Gyeongsan, Steinernema carpocapsae GSN1, S. glaseri Dongrae and S. longicaudum Nonsan strain) were infested bate insect, Galleria mellonella in log of dead pine tree. Result showed that, S. carpocapsae GSN1 strain has the highest pathogenicity. Heterorhabditis sp. Gyeongsan and S. carpocapsae GSN1 had infested bate insect, which located in 7.5 cm depth of pine log. EPN has pathogenicity against larva and adult of JPS. Spray application of EPNs against adult of JPS, Heterorhabditis sp. Gyeongsan strain showed higher pathogenicity than S. carpocapsae GSN1 strain. EPN infested larva of JPS was detected on bark spray treatment with S. carpocapsae GSN1 strain in dead pine log that naturally infested larva of JPS however, no EPN infested JPS was detected on soaking of pine log with suspension of S. carpocapsae GSN1 strain. Though EPN had found with low efficacy against JPS but still possible to apply to control the boring insect pest as they can control log dwelling insect.

• The Korean Journal of Pesticide Science
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• v.10 no.2
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• pp.107-116
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• 2006

Korean entomopathogenic nematode strain were evaluated against white grubs, Popillia quadriguttata, Ectinohoplia rufipes and Phyllopertha diversa in laboratory. In addition we examined the efficacy of entomopatogenic nematode, Heterorhabditis sp. KCTC 0991BP strain against white grubs in Seaside Golf Club in Gimpo, Gyeonggii and persistence of entomopatogenic nematode in fairway of Anyang Benest Golf Club in Gunpo, Gyeonggii. In laboratory Heterorhabditis bacteriophora Hamyang strain showed 95% mortality against Popillia quadriguttata. H. bacteriophora Hamyang strain and Heterorhabditis sp. KCTC 0991BP induced 80% mortality against 3rd instar of Ectinohoplia rufipes. However, the letter showed only 60% mortality against Phyllopertha diversa. White grub density was reduced to 39% by the application of Heterorhabditis sp. KCTC 0991BP at the rate of $2.24{\times}10^9$ infective juveniles $ha^{-1}$ compared with control in Seaside Golf Club. Heterorhabditis sp. KCTC 0991BP persisted for two months at $5{\sim}10$ cm soil depth but not persisted at $0{\sim}5$ cm soil depth for the same periods when applied at the rate of $2.47{\times}10^9$ infective juveniles $ha^{-1}$.

• Korean Journal of Organic Agriculture
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• v.23 no.4
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• pp.797-811
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• 2015

This study was carried out to identify the control effect of entomopathogenic microagent against Spodoptera exigua on organic chinese cabbage. In laboratory condition, insecticidal activity of 4 commercial BT pesticides against S. exigua were lower than 10% against second instar S. exigua. The insecticidal activity of entomopathogenic nematode were 33.3%, 83.3% and 100% at the concentration of $1{\times}10^2$, $3{\times}10^2$, $1{\times}10^3nematodes/ml$, respectively. Mixture of BT and nematode showed growth inhibition against S. exigua larvae. S. exigua nucleopolyhedrovirus (SeNPV) of $10^5PIBs/ml$ showed more than 70% insecticial activity. The yield of SeNPV was increased as in higher initial inoculation concentration of NPV, food supply, and growth temperature increased. In greenhouse experiment, the control value of BT and nematode mixture treatment was higher than BT and nematode treatment alone against S. exigua. In treatment of $10^7PIBs/ml$ of SeNPV, S. exigua was controlled completely. In farm condition, mixture of microbial agent and organic agricultural material showed higher control value against lepidopteran pest including S. exigua than BT single treatment.

• 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.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.906-909
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• 2001

In order to investigate fatty acid contents and effects of cell growth on the production of an extracellular protease and toxicity of exotoxin, several symbiotic bacteria with highly effective toxins were isolated from seven species of entomopathogenic nematodes belong in Steinernematidae(Steinernema glaseri XR-DR, S. glaseri XR-NC, S. glaseri XR-MK, S. carpocapsae XR-PC, S. maticola XR-MO, S. Longicaudum XR-LC) and Heterorhabditidae sp.(Heterorhabditis bacteriophora XR-HY). In the cell growth and exotoxin toxicity, XR-PC and XR-MK were superior to other species when cultured in vitro. The protease activity of XR-DR was remarkable compared to other species. In the case of XR-HY, the protease activity increased in parallel with cell growth. Interestingly the fatty acid contents of XR-PC and XR-HY were significantly different from those of other species 12:0, 14:0, 13:0 iso, 16:1 cis 5 and 17:0 cyclo.

• Journal of Microbiology and Biotechnology
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• v.21 no.8
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• pp.869-873
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• 2011

Most medium formulations for improving culture of entomopathogenic nematodes (EPN) based on protein sources have used enriched media like animal feed such as dried egg yolk, lactalbumin, and liver extract, among other ingredients. Most results, however, showed unstable yields and longer production time. Many of the results do not show the detailed parameters of fermentation. Soy flour, cotton seed flour, corn gluten meal, casein powder, soytone, peptone, casein hydrolysates, and lactalbumin hydrolysate as protein sources were tested to determine the source to support optimal symbiotic bacteria and nematode growth. The protein hydrolysates selected did not improve bacterial cell mass compared with the yeast extract control, but soy flour was the best, showing 75.1% recovery and producing more bacterial cell number ($1.4{\times}10^9$/ml) than all other sources. The highest yield ($1.85{\times}10^5$ IJs/ml), yield coefficient ($1.67{\times}10^6$ IJs/g medium), and productivity ($1.32{\times}10^7$ IJs/l/day) were also achieved at enriched medium with soybean protein.

• Journal of Microbiology and Biotechnology
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• v.28 no.1
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• pp.12-21
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• 2018

Photorhabdus temperata (PT), a gram-negative bacterium, lives symbiotically within entomopathogenic nematodes. The insecticidal compounds derived from Photorhabdus are used as biopesticides in agriculture. However, the physiological properties are not well characterized. In the course of our screening for neuroprotective and anti-neuroinflammatory substances from natural products, the culture broth of PT showed considerable activities. By activity-guided purification, five anthraquinones, namely, 3-methoxychrysazine (1), 1,3-dimethoxy-8-hydroxy-9,10-anthraquinone (2), 1,3,8-trihydroxy-9,10-anthraquinone (3), 3,8-dihydroxy-1-methoxy-9,10-anthraquinone (4), and 1,3,4-trimethoxy-8-hydroxy-9,10-anthraquinone (5), were isolated from the ethyl acetate fraction of the PT culture broth. Among the isolated compounds, $75{\mu}M$ 3 significantly protected mouse hippocampal neuronal cells (HT22) against 5 mM glutamate-induced cell death via the inhibition of reactive oxygen species production, $Ca^{2+}$ influx, and lipid peroxidation. Additionally, 3 and 4 effectively suppressed the interferon-${\gamma}$-induced neuroinflammation of mouse-derived microglial cells (BV2) at 10 ng/ml, via the reduction of nitric oxide, interleukin-6, and tumor necrosis factor-${\alpha}$. Anthraquinones 3 and 4 derived from the PT culture broth are a potential starting point to discover neuroprotective and anti-neuroinflammatory drug leads. The novel compound 5 is reported for the first time in this study.