• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.2
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• pp.141-145
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• 2000

Entomopathogenic nematodes are being used for insect control. We purified a toxin secreted by the insect-pathogenic bacterium, Xenorhadbus nematophilus, which lives in the gut of entomopathogenic nematodes. Culture broth of X. nematophilus was separated by centrifugation and concentrated by ultration. The concentrated culture broth was applied to a DEAE Sephadex A-50 column, and proteins were eluted stepwise with increasing concentrations of KCI. Fractions column. The molecty weight of purified toxin was39 kDa on SDS-PAGE, and Fourier tranformed infrared (FTIR) spectroscopy indicated that this toxin could be a new protein exhiting the charactristics of C=O stretching peak near 1650cm-1.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.5
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• pp.379-382
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• 2004

Entomopathogenic nematodes are used for insect control. Herein, an extracellular protease was partially purified from a culture supernatant of Xenorhabdus nematophilus, a symbiotic bacterium of an entomopathogenic nematode, Steinernema glaseri: using precipitation with $80\%$ v/v isopropyl alcohol followed by gel permeation chromatography with a packed Sephacryl S-300 HR media. The partially purified protease exhibited maximal activity at pH 7 in the presence of 1 mM $CaCl_2$. The protease was identified as a metallo-protease based on the inhibition of its activity by the metal chelating agent, EDTA.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.117-123
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• 2010

BT toxin is produced by a soil bacterium Bacillus thuringiensis and has long been used as a biological insecticide without any competition. Recently, Photorhabdus, a symbiotic bacterium from entomopathogenic nematodes, family Heterorhabditae, has been researched and discussed as alternatives to B. thuringiensis. Photorhabdus, which lives in the gut of entomopathogenic nematodes, is a highly virulent pathogen of a wide range of insect larvae. When an insect is infected by the nematodes, the bacteria are released into the cadaver, and produce a number of insecticidal toxins. The biological role of the different Photorhabdus toxins in the infection process is still unclear. Photorhabdus toxin complex (Tc) is highly secreted gut-active toxin and has been characterized as a potent three-component (A, B and C) insecticidal protein complex. These components are necessary for full oral activity against insect larvae. The Photorhabdus PirAB binary toxins exhibit a potent injectable activity for Galleria mellonella larvae, and have oral toxicity against mosquitoes and caterpillar pest Plutella xylostella. Other toxin, 'makes caterpillars floppy' (Mcf) showed injectable activity on caterpillars. Recombinant Mcf triggers apoptosis in both insect hemocytes and the midgut epithelium and carries a BH3 domain. In this review, the relationship between the Photorhabdus and the nematode is discussed and recent important insecticidal toxins from Photorhabdus are described.

• Biotechnology and Bioprocess Engineering:BBE
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• v.4 no.1
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• pp.12-16
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• 1999

Xenorhabdus nematophilus sp., an insect-pathogenic bacterium, was newly isolated from Korean entomopathogenic nematode of Steinernema carpocapsae, which can be used as a useful bioinsecticide. Primary and secondary form variants of Xenorhabdus nematophilus were observed when cultured in vitro. Primary form variants adsorbed bromothymol blue, while secondary form did not. However, many other characters of two variants were very similar. The variants were all rod-shaped and cell size was highly variable ranging from 0.5 by 2.0 ${\mu}$m to 1.0 by 5.0 ${\mu}$m. Both produced highly toxic substances and killed the insect larva within 20∼38 hr, indicating that insect pathogenicity of Xenorhabdus is not directly associated with its phase variation. In addition, cell-free culture supernatant of Xenorhabdus was sufficient to kill the insect larva by injecting it ito insect hemolymph; however, cell-harboring culture broth was more effective for killing the insect. The use of Xenorhabdus nematophilus may provide a potential alternative to Bacillus thuringiensis (Bt) toxins.

• KSBB Journal
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• v.14 no.3
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• pp.303-308
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• 1999

An in vitro culture method for entomopathogenic nematode Steinernema glaseri was developed. A symbiotic bacterium was isolated from Steinernema glaseri and identified as Xenorhabdus nematophilus. Phase variation that differed in some biochemical characteristics of symbiotic bacterium was observed. Entomopathogenic nematodes carried only phase I bacterium in their guts. Phase I bacterium could be converted into phase II form in in vitro culture medium consisting of 5% yeast extract, 0.5% NaCl, 0.05% $K_2HPO_4$, $0.02% MgSO_4$.$7H_2O$. The optimum temperature for bacterial growth was $28^{\circ}C$. The pH of the culture medium increased up to 9.0-9.5 during the exponential growth period of the culture, regardless of initial pH 6-7. Various culture media such as chicken offal, dog food, bovine liver, peanut, and so on were tested for in vitro culture of the nematodes. The best medium for Steinernema glaseri production was obtained from concentrated homogenate of bovine liver and the nematode growth was highest at 80% bovine liver. In the co-culture of entomopathogenic nematode with its symbiont, the growth rate of nematodes was 2 times faster than that without its symbiont and the nematode concentration reached about $5.5\times10^4$/mL within 15 days.

• Journal of Microbiology and Biotechnology
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• v.17 no.3
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• pp.420-427
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• 2007

We investigated the temperature effects on the virulence, development, reproduction, and otility of two Korean isolates of entomopathogenic nematodes, Steinernema glaseri Dongrae strain and S. longicaudum Nonsan strain. In addition, we studied the growth and virulence of their respective symbiotic bacterium, Xenorhabdus poinarii for S. glaseri and Xenorhabdus sp. for S. longicaudum, in an insect host at different temperatures. Insects infected with the nematode-bacterium complex or the symbiotic bacterium was placed at $13^{\circ}C,\;18^{\circ}C,\;24^{\circ}C,\;30^{\circ}C,\;or\;35^{\circ}C$ in the dark and the various parameters were monitored. Both nematode species caused mortality at all temperatures tested, with higher mortalities occurring at temperatures between $24^{\circ}C\;and\;30^{\circ}C$. However, S. longicaudum was better adapted to cold temperatures and caused higher mortality at $18^{\circ}C$ than S. glaseri. Both nematode species developed to adult at all temperatures, but no progeny production occurred at $13^{\circ}C\;or\;35^{\circ}C$. For S. glaseri, nematode progeny production was best at inocula levels above 20 infective juveniles/host at $24^{\circ}C\;and\;30^{\circ}C$, but for S. longicaudum, progeny production was generally better at $24^{\circ}C$. Steinernema glaseri showed the greatest motility at $30^{\circ}C$, whereas S. longicaudum showed good motility at $24^{\circ}C\;and\;30^{\circ}C$. Both bacterial species grew at all tested temperatures, but Xenorhabdus sp. was more virulent at low temperatures $(13^{\circ}C\;and\;18^{\circ}C)$ than X. poinarii.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2001.09a
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• pp.107-110
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• 2001

The biological control potential of entomopathogenic nematodes (EPN) can be enhanced by improved culture efficiency. Optimization of media is a key factor for improving in vitro mass production of entomopathogenic nematodes. EPN yield was dependant of complex medium concentration, of which mixture is carbohydrates, lipids, proteins, salts, and growth factors, on the growth of Heterorhabditis bacteriophora and its symbiotic bacterium Photorhabdus luminescensLipids.

• Journal of Microbiology and Biotechnology
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• v.11 no.4
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• pp.644-648
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• 2001

The biological control potential of entomopathogenic nematodes (EPN) can be enhanced by improved culture efficiency. Optimization of the media is a key factor for improving in vitro mass production of entomopathogenic nematodes. This study reports the effect of medium concentration. The medium is a combination of carbohydrates, lipids, proteins, sats, and growth factors, on the growth of Heterorhabditis bacteriophora and its symbiotic bacterium Photorhabdus liminescens. The overall optimal medium concentration for nematode recovery, hermaphrodite size, bacterial mass, infective juveniles (IJs) yield, and doubling time was 84 g/l. At this concentration rate, the doubling time of IJs production and the biomass of symbiotic bacteria was 1.6 days and 12.8 g/l, respectively. The maximum yield of $2.4{\times}{10^5}IJs/ml$ was attained within a one-generation cycle (eight days). The yield coefficient was $2.8{\times}{10^6}$ IJs/g medium, and the maximum productivity was $3.1{\times}{10^7}$ IJs per day. Medium concentration affected two independent factors, recovery and hermaphrodite size, which in turn influenced the final yield.

• Korean journal of applied entomology
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• v.51 no.1
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• pp.39-45
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• 2012

The entomopathogenic bacterium, $Xenorhabdus$ sp., was isolated from an entomopathogenic nematode, $Steinernema$ $monticolum$. When these bacteria were injected into the hemocoel of the diamondback moth, $Plutella$ $xylostella$, they caused significant mortality. However, the bacterium was not pathogenic when it was administered orally. This study showed that $Xenorhabdus$ sp. significantly enhanced oral pathogenicity of $Bacillus$ $thuringiensis$ (Bt) against the last instar larvae of $P.$ $xylostella$. Different ratios of culture broth of $Xenorhabdus$ sp. and Bt showed significantly different pathogenicities against $P.$ $xylostella$. In field tests, the optimal bacterial mixture significantly enhanced control efficacy against $P.$ $xylostella$ compared to Bt treatment alone. These results demonstrated that $Xenorhabdus$ sp. culture broth can be developed as a potent biopesticide by enhancing the insecticidal efficacy of Bt.

• KSBB Journal
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• v.14 no.2
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• pp.198-204
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• 1999

Asymbiotic bacterium with highly effective toxins was isolated from entomopathogenic nematode Steinernema glaseri which has been widely used against various soil-inhabiting pests. The symbiont of S. glaseri was identified as Xenorhabdus nematophilus sp. by using several biochemical and physiological tests. When this strain was released into the hemolymph of insect larva, it produced highly toxic substances and killed the larva within 2 days. Two colony forms that differed n some biochemical characteristics were observed when cultures in vitro. Phase l colonies were mucid and difficult to be dispersed in liquid. Phase II was not mucoid and was easily dispersed in liquid. It did not adsorb neutral red or bromothymol blue. Rod-shaped cell size was highly variable between two phases, ranging 2-10 ${\mu}{\textrm}{m}$. It was also found that only infective-stage nematodes can carry only primary-phase Xenorhabdus in their intestine.