• The Korean Journal of Pesticide Science
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• v.3 no.3
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• pp.60-67
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• 1999

Insecticidal activities of imidacloprid to sucking type insect pests, brown planthopper (BPH, Nilaparvata lugens) and peen peach aphid (GPA Myzus persicae), to chewing type insect pests, tobacco cut worm (TCW; Spodoptera litura) and beet armyworm (BAW, Spodoptera exigua) and to spiders, Pirata subpiraticus, Pachygnata clercki and Ummeliata insecticeps, as natural enemies were investigated by several bioassay methods. $LD_{50}$ of the chemical by topical treatment to BPH was 0.015 ${\mu}g/g$(48 hrs), while $LC_{50}s$ by leaf dipping and root zone treatment were 18.1 and 21.5 ppm, respectively. There was no difference in insecticidal activities between leaf dipping and root zone treatment. Imidacloprid also showed ovicidal effect of root zone treatment and its $LC_{50}$ was 6.8ppm $LD_{50}$ (48 hrs after treatment) of imidacloprid to GPA was $0.4{\mu}g/g$ in case of topical application and $LC_{50}s$ by leaf dipping and root zone treatment were 1.9 ppm and 13.7 ppm respectively. Leaf dipping was more effective than root zone treatment in GPA At topical application $LD_{50}$ (48 hrs after treatment) of imidacloprid to chewing type insect pests, TCW and BAW, were greater than 1,492 and $312{\mu}g/g$ and $LC_{50}s$ by leaf dipping method were 4,803 and Heater than 5,000ppm respectively. This means that imidacloprid has much less effect on chewing type insect pests, TCW and BAW. $LD_{50}$(48 hrs after treatment) of imidacloprid to wandering spiders, Pirata subpiraticus, Pachygnata clercki at topical application were greater dan $2941{\mu}g/g$ and greater than $2,976{\mu}g/g$ respectively and that to webbing spider, Ummeliata insecticeps, was 357 ${\mu}g/g$. Imidacloprid showed very low toxicity to the spiders and its selective toxicity ratios between spiders and BPH were greater than 19,600, greater than 19,800 and 23,800, respectively.

• 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.

• KSBB Journal
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• v.15 no.1
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• pp.100-105
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• 2000

Optimal medium composition, culture conditions, characteristics of phase variation and activity of insecticidal toxin by Xenorhabdus nematophilus isolated and identified from Korean entomopathogenic nematode Steinernema carpocapsae were examined. Optimal medium composition of this strain was 50-70 g/L yeast extract, 3 g/L $K_{2}HPO_{4}$, 1g/L $NH_{4}H_{2}PO_{4}$, 2g/L ${MgSO}_4$$\cdot$${7H}_{2}O$, 10g/L NaCl and, these, yeast extract was found as a limiting nutrient for cell growth. When Monod equation was applied, maxmum specific growth rate and Monod constant were estimated as 0.13 $hr^{-1}$ and 20g/L, respectively. The pH of culture medium increased up to 8.5-9.5 regardless of initial pH 6-7 as the cells continued to grow. The specific growth rate in a 7 L fermentor was 0.18 $hr^{-1}$, which was enhancement 1.4 fold compared to a flask culture. In case of phase variation, phase I fraction was maintained above 90% at the stationary phase for both flask and fermentor cultures. According to oral toxicity test of Gallena mellonella by Xenorhabdus nematophilus, the addition of cell pellets into feed inhibited normal growth of insect larvae and killed completely then after 20 days cultivation. When culture supernatant of this strain was injected into hemolymph of insect larva, the toxicity was strongest at 24hr cultivation in the early exponential phase and gradually decreased as the culture time proceeded.

• BMB Reports
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• v.40 no.5
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• pp.773-782
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• 2007

Different isolates of the soil bacterium Bacillus thuringiensis produce multiple crystal (Cry) proteins toxic to a variety of insects, nematodes and protozoans. These insecticidal Cry toxins are known to be active against specific insect orders, being harmless to mammals, birds, amphibians, and reptiles. Due to these characteristics, genes encoding several Cry toxins have been engineered in order to be expressed by a variety of crop plants to control insectpests. The cotton boll weevil, Anthonomus grandis, and the fall armyworm, Spodoptera frugiperda, are the major economically devastating pests of cotton crop in Brazil, causing severe losses, mainly due to their endophytic habit, which results in damages to the cotton boll and floral bud structures. A cry1Ia-type gene, designated cry1Ia12, was isolated and cloned from the Bt S811 strain. Nucleotide sequencing of the cry1Ia12 gene revealed an open reading frame of 2160 bp, encoding a protein of 719 amino acid residues in length, with a predicted molecular mass of 81 kDa. The amino acid sequence of Cry1Ia12 is 99% identical to the known Cry1Ia proteins and differs from them only in one or two amino acid residues positioned along the three domains involved in the insecticidal activity of the toxin. The recombinant Cry1Ia12 protein, corresponding to the cry1Ia12 gene expressed in Escherichia coli cells, showed moderate toxicity towards first instar larvae of both cotton boll weevil and fall armyworm. The highest concentration of the recombinant Cry1Ia12 tested to achieve the maximum toxicities against cotton boll weevil larvae and fall armyworm larvae were 230 ${\mu}g/mL$ and 5 ${\mu}g/mL$, respectively. The herein demonstrated insecticidal activity of the recombinant Cry1Ia12 toxin against cotton boll weevil and fall armyworm larvae opens promising perspectives for the genetic engineering of cotton crop resistant to both these devastating pests in Brazil.

• The Korean Journal of Pesticide Science
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• v.12 no.3
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• pp.262-269
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• 2008

This study was performed to investigate the toxicity of 39 registered insecticides to the susceptibility, systemic effect, and residual effect and control effect against Pine sawyer beetle, Monochamus saltuarius. Eleven kinds of chemicals such as fenitrothion, fenthion, phenthoate, phosphamidon, dinotefuran, actamiprid, clothianidin, thiacloprid, thiamethoxam, esfenvalerate+fenitrothion, and fipronil were showed 100% insecticidal activity both in body spray and twig dipping bioassay. Among these chemicals, fenitrothion and fenthion were showed 100% insecticidal activity when sprayed at 4000 times diluted solutions, and phenthoate, thiacloprid, thiamethoxam and fipronil were showed 100% insecticidal activity when sprayed at 2000 times diluted solution. Root systemic effect was showed 100% mortality in phosphamidon, clothianidin, thiamethoxam, and 77.7% in thiacloprid. In residual effect, fenitrothion and thiamethoxam were showed 80% mortality fifteen days after treatment (DAT), and fenthion, phosphamidon, clothianidin were showed 80% mortality ten DAT, fenitrothion, thiamethoxam, fipronil showed 100% mortality in seven DAT, thiacloprid was showed 100% mortality in three DAT. Fenthion and phenthoate were showed 100% mortality one DAT. In the control effect, 6 kinds of chemicals were showed 100% mortality one DAT and all chemicals showed 100% mortality three DAT.

• Korean journal of applied entomology
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• v.45 no.3 s.144
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• pp.357-362
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• 2006

Bacillus thuringiensis strains were isolated from the domestic soil and a strain was selected that had a new host range and high toxicity against agriculture insect pest. The 142 samples of soil were sampled from the mountains, paddy fields and patches, in Daejon, Chungnam, Chungbuk and Jeonbuk and used for the investigation. Sixteen B. t strains were isolated from 12 samples among collected samples. There were 11 strains that showed toxical activity on Plutella xylostella (Lepidoptera: Yponomeutidae), 7 steins on Spodoptera litura (Lepidoptera: Noctuidae), 5 strains on Arete coerulea (Lepidoptera: Noctuidae), 5 strains on Culex pipiens pallens (Diptera: Culicidae) among the 16 isolated B. t strains. But there were not any strains that showed activity against Hyphanria cunea (Lepidoptera: Arctiidae) and Sitophilus oryzae (Coleoptera: Rhynchophoridae). And also some of B. thuringiensis strains showed insecticidal activity with 2, 3 or 4 kinds of insects. But there were also 3 strains that did not show any activities to the 6 insects which were used in the experiment. When examined with a phase-contrast microscope, the insecticidal crystal protein produced from 16 selected strains had 13 bipyramidal and 3 spherical shapes. The insecticidal bioactivity of the S. litura showed 100% mortality when there were $1.3{\times}10^{7}\;(cfu/ml)$ of CAB109 isolates.

• Korean Journal of Environmental Biology
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• v.34 no.3
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• pp.208-211
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• 2016

Essential oils (EOs) extracted from plants possess various biological activities and have been considered as natural insecticides due to their potent insecticidal activities. In regard to develop natural insecticides, EOs are formulated as an emulsifiable concentrate and their acute toxicity against to fishes were determined in a static condition using Cyprinus carpio. Coriander EO was used as an active ingredient mixed with ethanol for solvent and various surface active agents. The tested EOs were obtained from a commercial market, and three different extractions were also undertaken to produce EO using steam distillation, solvent extraction, and supercritical fluid extraction. Among the emulsifiable concentrate including a commercial coriander EO, surface active agents such as Tween 80, sodium dodecyl benzene sulfonate (SDBS), and mixture of SDBS and Nonidet showed acute toxicity to the fish. With the three different EO extraction, coriander EO obtained from supercritical fluids with Triton X-100 exhibited acute toxicity to C. carpio. Taken together, Tetgitol and Nondet are considered as surface active agents for the emulsifiable formulation of coriander EO.

• BMB Reports
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• v.36 no.3
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• pp.294-298
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• 2003

The pathological effect of the Bacillus thuringiensis Cry $\delta$-endotoxins on susceptible insect larvae had extensive damage on the midgut epithelial cells. In this study, an ex vivo assay was devised for assessing the insecticidal potency of the cloned Cry4B mosquito-larvicidal protein that is expressed in Escherichia coli. Determination of toxicity was carried out by using a cell viability assay on the midguts that were dissected from 5-day old Aedes aegypti mosquito larvae. After incubation with the toxin proteins, the number of viable epithelial cells was determined photometrically by monitoring the quantity of the bioreduced formazan product at 490 nm. The results showed that the 65-kDa trypsin-activated Cry4B toxin exhibited toxic potency ca. 3.5 times higher than the 130-kDa Cry4B protoxin. However, the trypsin-treated products of the non-bioactive Cry4B mutant (R158A) and the lepidopteran-specific Cry1Aa toxin displayed relatively no ex vivo activity on the mosquito-larval midguts. The ex vivo cytotoxicity studies presented here confirms data that was obtained in bioassays.

• Journal of Microbiology and Biotechnology
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• v.17 no.9
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• pp.1498-1503
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• 2007

To identify novel crystal proteins, Bacillus thuringiensis 2385-1 was isolated from Korean soil samples and characterized. The H-serotype of 2385-1 was identical to that of subsp. kenyae (H4a4c), and its crystal toxin was bipyramidal-shaped. However, 2385-1 showed a much higher toxicity towards Plutella xylostella and Spodoptera exigua larvae than subsp. kenyae. In addition, the crystal protein profile and plasmid DNA pattern of 2385-1 differed from those of subsp. kenyae. To verify the crystal protein gene types of 2385-1, a PCR-RFLP analysis was performed, and the results revealed that 2385-1 contained two novel cry1-type crystal protein genes, cryl-5 and cry1-12, in addition to the crylJal gene. The deduced amino acid sequences of cryl-5 and cry1-12 showed a 97.9% and 75.7% sequence similarity with the CrylAb and CrylJa crystal proteins, respectively. Among the novel crystal proteins, Cry1-5 showed a high toxicity towards P. xylostella and S. exigua larvae. In conclusion, B. thuringiensis 2385-1 is a new isolate in terms of its gene types, and should be a promising source for an insecticide to control lepidopteran larvae.

• The Korean Journal of Pesticide Science
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• v.10 no.3
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• pp.218-224
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• 2006

Toxicities of 34 agricultural insecticides was tested against Monomorium pharaonis workers by diet dipping method. Ten insecticides among them, bifenthrin, chlorpyrifos, chlorpyrifos-methyl, cypermethrin, dichlorvos, fenthion, fenitrothion, methidathion, pirimifos-methyl, and phenthoate showed 100% rapid mortality. $LC_{50}$ (ppm) values of selected insecticides were appeared on the order of pirimifos-methyl (0.33), chlorpyrifos-methyl (0.76), phenthoate (1.70), bifenthrin (1.78), dichlorvos (2.50), cypermethrin (9.92), chlorpyrifos (22.21), fenitrothion (36.58), fenthion (40.96), and methidathion (64.34). $LT_{50}$ (day) values by diet dipping method showed that dichlorvos, benfuracarb and cypermethrin acted more rapid than boric acid and hydramethylnon. The values of the former three were 0.25, 0.38 and 0.27 days, and those of the latter two were 3.4 and 2.6 days, respectively. In persistance effect tests, chlorpyrifos-methyl, fenthion and methidathion showed over 90% insecticidal activity for 13 days.