• Journal of Microbiology and Biotechnology
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• v.23 no.8
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• pp.1107-1115
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• 2013

The Cyt1Aa protein of Bacillus thuringiensis susbp. israelensis elaborates demonstrable toxicity to mosquito larvae, but more importantly, it enhances the larvicidal activity of this species Cry proteins (Cry11Aa, Cry4Aa, and Cry4Ba) and delays the phenotypic expression of resistance to these that has evolved in Culex quinquefasciatus. It is also known that Cyt1Aa, which is highly lipophilic, synergizes Cry11Aa by functioning as a surrogate membrane-bound receptor for the latter protein. Little is known, however, about whether Cyt1Aa can interact similarly with other Cry proteins not primarily mosquitocidal; for example, Cry2Aa, which is active against lepidopteran larvae, but essentially inactive or has very low toxicity to mosquito larvae. Here we demonstrate by ligand binding and enzyme-linked immunosorbent assays that Cyt1Aa and Cry2Aa form intermolecular complexes in vitro, and in addition show that Cyt1Aa facilitates binding of Cry2Aa throughout the midgut of C. quinquefasciatus larvae. As Cry2Aa and Cry11Aa share structural similarity in domain II, the interaction between Cyt1Aa and Cry2Aa could be a result of a similar mechanism previously proposed for Cry11Aa and Cyt1Aa. Finally, despite the observed interaction between Cry2Aa and Cyt1Aa, only a 2-fold enhancement in toxicity resulted against C. quinquefasciatus. Regardless, our results suggest that Cry2Aa could be a useful component of mosquitocidal endotoxin complements being developed for recombinant strains of B. thuringiensis subsp. israelensis and B. sphaericus aimed at improving the efficacy of commercial products and avoiding resistance.

• BMB Reports
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• v.47 no.10
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• pp.546-551
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• 2014

The insecticidal activity of Bacillus thuringiensis (Bt) Cry toxins involves toxin stabilization, oligomerization, passage across the peritrophic membrane (PM), binding to midgut receptors and pore-formation. The residues Arg-158 and Tyr-170 have been shown to be crucial for the toxicity of Bt Cry4Ba. We characterized the biological function of these residues. In mosquito larvae, the mutants R158A/E/Q (R158) could hardly penetrate the PM due to a significantly reduced ability to alter PM permeability; the mutant Y170A, however, could pass through the PM, but degraded in the space between the PM and the midgut epithelium. Further characterization by oligomerization demonstrated that Arg-158 mutants failed to form correctly sized high-molecular weight oligomers. This is the first report that Arg-158 plays a role in the formation of Cry4Ba oligomers, which are essential for toxin passage across the PM. Tyr-170, meanwhile, is involved in toxin stabilization in the toxic mechanism of Cry4Ba in mosquito larvae.

• The Korean Journal of Pesticide Science
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• v.3 no.1
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• pp.46-50
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• 1999

Methanol extracts of 9 kinds of native medicinal plants(Taraxacum platycarpum, leaf; Pinus densiflora, leaf; Artemisia prinseps, leaf; Allium tuberosum, leaf; Cassia obtussifolia, whole; Sophora angestifolia, root; Stemonae sessilifolia, root; Lonicera japonica stem, leaf, flower; and Clivia miniata) were investigated for insecticidal effect. Methanol extracts of Pinus densiflora leaves and Sophora angestifolia radix showed relatively good insecticidal activity against Culex pipiens pallens larvae. Strong larvicidal activity against the Musca domestica larvae was produced from methanol extracts of Taraxacum platycarpum leaves and Allium tuberosom leaves. while Stemonae radix showed moderate activity. All materials tested revealed little or weak insecticidal activity against M. domestica adults.

• Korean journal of applied entomology
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• v.43 no.2
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• pp.169-173
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• 2004

Control effects of some insecticides were evaluated against the stone leek leafminer, Liriomyra chinensis Kato (Diptera： Agromyzidae) with the some different treatment methods. Insecticidal activities effects were estimated on the different development stages of the insects on welsh onion. The insecticides that controlled L. chinensis eggs with over 83％ efficacy were spinosad, dimethoate, emamectin, and cartap. The insecticides that showed over 87％ of larvicidal activity were dimethoate and cartap. Dimethoate showed 93.3％ insecticide residual activity for 3 days a(ter treatment as a foliar spray. For control of pupae, the insecticides that showed over 88％ of contact insecticidal activity were terbufos GR and cartap GR. Both dimethoate and cartap had high adulticidal activity with over 95％ control efficacy.

• BMB Reports
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• v.39 no.3
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• pp.270-277
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• 2006

Helices 4 and 5 of the Bacillus thuringiensis Cry4Ba $\delta$-endotoxin have been shown to be important determinants for mosquito-larvicidal activity, likely being involved in membrane-pore formation. In this study, the Cry4Ba mutant protein containing an additional engineered tryptic cleavage site was used to produce the $\alpha4$-$\alpha5$ hairpin peptide by an efficient alternative strategy. Upon solubilization of toxin inclusions expressed in Escherichia coli and subsequent digestion with trypsin, the 130-kDa mutant protoxin was processed to protease-resistant fragments of ca. 47, 10 and 7 kDa. The 7-kDa fragment was identified as the $\alpha4$-loop-$\alpha5$ hairpin via N-terminal sequencing and mass spectrometry, and was successfully purified by size-exclusion FPLC and reversed-phase HPLC. Using circular dichroism spectroscopy, the 7-kDa peptide was found to exist predominantly as an $\alpha$-helical structure. Membrane perturbation studies by using fluorimetric calcein-release assays revealed that the 7-kDa helical hairpin is highly active against unilamellar liposomes compared with the 65-kDa activated full-length toxin. These results directly support the role of the $\alpha4$-loop-$\alpha5$ hairpin in membrane perturbation and pore formation of the full-length Cry4Ba toxin.

• BMB Reports
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• v.34 no.2
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• pp.130-133
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• 2001

The amino acid sequences of 130 kDa and 72 kDa proteins responsible for the larvicidal activity of Bacillus thuringiensis var israelensis (Bti) were analyzed by hydrophobic moment plots. A search for highly amphiphilic $\alpha$-helices was made in these proteins using the helical hydrophobic moment as a criterion of amphiphilicity The protein segments of the largest hydrophobic moments were analyzed. In the present communication we report the surface seeking helices in 130 kDa and 72 kDa proteins of Bacillus thuringiensis var israelensis. It is assumed that the surface seeking segments may participate in one of the membrane-related functions of Bacillus thuringiensis.

• The Korean Journal of Pesticide Science
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• v.6 no.4
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• pp.257-263
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• 2002

This study was carried out not only to investigate the toxicities of 12 registered insecticides on different developmental stages, but also to determine the sublethal effects on longevity and reproduction of newly emerged adult female and development of the next generation in the rice skipper, Parnara guttata. Fenitrothion, fenthion, cartap hydrochloride, ethofenprox highly suppressed egg-hatch. All insecticides treated showed high larvicidal activity on the 1st to 2nd instar larva. The insecticides showed higher larvicidal activities on the 5th instar larva were fenitrothion, fenthion, ethofenprox, fipronil, methoxyfenozide, tebufenozide and Bt. var. kurstaki. The sublethal doses of fenthion, tebufenozide, cartap hydrochloride, methoxyfenozide, ethofenprox, imidacloprid and fipronil shortened the longevities of newly emerged adult female from the treated larva ($3{\sim}4$ instar). BPMC, imidacloprid, ethofenprox, fipronil and methoxyfenozide delayed the preoviposition periods of adult females and decreased the number of eggs laid when they were treated at the larval stages of the previous generation. Ethofenprox caused severe sublethal effects on P. guttata offspring, completely blocking the production. All insecticides except fenitrothion affected the egg viability, and all eggs from the adult females emerged from the survivors treated larvae with imidacloprid or fipronil fail to hatch. IGRs, methoxyfenozide and tebufenozide showed an adverse effect on the development of next generation larva.

• The Korean Journal of Pesticide Science
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• v.4 no.3
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• pp.75-81
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• 2000

These studies were carried out to investigate the toxicities of 38 registered insecticides to the sweetpotato whitefly (Trialeurodes vaporariorum). Insecticide activities were evaluated by testing systemic action and residual effect in the laboratory, and control efficacy in the greenhouse. All experiments were tested at the recommended concentration(ppm) of each insecticides. Insect growth regulators (IGRs), only pyriproxyfen showed over 90% of ovicidal effect. The insecticides that showed over 90% of larvicidal activity oil 3rd nymphal instars were abamectin, acetamiprid, chlorpyrifos-methyl, imidacloprid, pyripoxyfen, and acetamiprid+ethofenprox. Insecticides with 100% adulticidal activity were abamectin, acephate, acetamiprid, benfurcarb, bifenthrin, furathiocarb, endosulfan, fenitrothion, imidacloprid, phenthoate, pymetrozine, acetamiprid + ethofenprox, ethofenprox + diazinon, furathiocarb + difluberlzuron, and triazamate+${\alpha}$-cypermethrin. Abamectin, acetamiprid, imidacloprid, pyriproxyfen, and acetamiprid + ethofenpox showed both residual effect and systemic activity. In tile control efficacy test on B. tabaci, 90% control values were obtained at 11th day after treatment of the insecticides including abamectin, acetamiprid, imidacloprid, pyripoxyfen and acetamiprid + ethofenprox. These results indicate that abamectin, acetamiprid, imidacloprid, pyriproxyfen and acetamiprid + ethofenprox can be used for tile control of B. tabaci in field.

• BMB Reports
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• v.34 no.4
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• pp.293-298
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• 2001

Tryptic activation of the 130-kDa Bacillus thuringiensis Cry4B $\delta$-endotoxin produced protease-resistant products of ca. 47 kDa and ca. 21 kDa. The 21-kDa fragment was identified as the N-terminal five-helix bundle (${\alpha}1-{\alpha}5$,) which is a potential candidate for membrane insertion and pore formation. In this study, we constructed the recombinant clone over-expressing this putative pore-forming (PPF) fragment as inclusion bodies in Escherichia coli. The partially purified inclusions were composed of a 23-kDa protein, which cross-reacted with Cry4B antibodies, and whose N-terminus was identical to that of the 130-kDa protein. Dissimilar to protoxin inclusions, the PPF inclusions were only soluble when the carbonate buffer, pH 9.0, was supplemented with 6 M urea. After renaturation via a stepwise dialysis, the refolded PPF protein appeared to exist as an oligomer and was structurally stable upon trypsin treatment. Unlike the 130kDa protoxin, the refolded protein was able to release entrapped glucose from liposomes, and showed comparable activity to the full-length activated toxin, although it lacks larvicidal activity These results, therefore, support the notion that the PPF fragment that consists of ${\alpha}1-{\alpha}5$ of the activated Cry4B toxin is involved in membrane pore-formation.

• The Korean Journal of Pesticide Science
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• v.11 no.1
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• pp.52-58
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• 2007

ALA (5-aminolevulinic acid) has been proposed as a tetrapyrrole-dependent photodynamic herbicide and insecticide by the action of the protoporphyrinogen IX oxidase (Protox IX). The present study was conducted to determine growth responses of plant and insects to ALA, biodegradable biopesticidal substance. In the paddy condition experiment, plant height and shoot fresh weight of barnyardgrass (Echinochloa crus-galli) was more reduced by ALA than rice plants, even though both plant species show great phytotoxicity. Hairy crabgrass (Digitaria sanguinalis), a monocot weed, was more sensitive to ALA at 5mM under upland condition when ALA applied on the foliage, compared with soybean (Glycine max) as a dicot crop. ALA solutions were tested for their insecticidal and larvicidal activities against Spodaptera exigua (Hubner) and Tetranychus urticae Koch. by foliar application and leaf-dipping method. The result showed higher insecticidal activity of ALA at 10mM and its mixture with insecticide luferon against S. exigua. Strongest insecticidal activity against T. urticae was observed from the ALA solution at 10mM 72 days after application. This results show that ALA solution had potent herbicidal and insecticidal activities against agricultural pests even though their activities were lower than those of synthetic pesticides.