Identification and characterization of proteins from Bacillus thuringiensis with high toxic activity against the sheep blowfly, Lucilia cuprina

Gough, J.; Kemp, D. H.; Akhurst, Raymond J.; Pearson, Roger; Kongsuwan, Kritaya

doi:10.1016/j.jip.2005.05.012

Cited by 18 publications

(26 citation statements)

References 25 publications

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“…The observed lethality is attributed to Cry1Ab21, as we have shown that this strain does not produce detectable levels of ␤-exotoxin (8). It is difficult to directly compare this level of toxicity to the levels of toxicity of other isolates and strains of B. thuringiensis that have been described previously because the methods used by other investigators vary.…”

Section: Discussionmentioning

confidence: 86%

“…To demonstrate that the toxicity of IS5056 against T. ni larvae was not due to ␤-exotoxin, bioassays were performed as described above using autoclaved crystal-spore suspensions (121°C for 20 min) at a concentration of 400 g/ml. This procedure has been shown to inactivate protein crystal toxins but not ␤-exotoxin (8). In addition to ␤-exotoxin inactivation, the effect of autoclaving on the shape of the crystalline inclusion of IS5056 was observed by contrast field microscopy at a magnification of ϫ1,000.…”

Section: Methodsmentioning

confidence: 99%

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Novel Isolate of Bacillus thuringiensis subsp. thuringiensis That Produces a Quasicuboidal Crystal of Cry1Ab21 Toxic to Larvae of Trichoplusia ni

Święcicka

Bideshi

Federici

2008

Appl Environ Microbiol

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A new isolate (IS5056) of Bacillus thuringiensis subsp. thuringiensis that produces a novel variant of Cry1Ab, Cry1Ab21, was isolated from soil collected in northeastern Poland. Cry1Ab21 was composed of 1,155 amino acids and had a molecular mass of 130.5 kDa, and a single copy of the gene coding for this endotoxin was located on a ϳ75-kbp plasmid. When synthesized by the wild-type strain, Cry1Ab21 produced a unique, irregular, bipyramidal crystal whose long and short axes were both approximately 1 m long, which gave it a cuboidal appearance in wet mount preparations. In diet incorporation bioassays, the 50% lethal concentrations of the crystal-spore complex were 16.9 and 29.7 g ml ؊1 for second-and fourth-instar larvae of the cabbage looper, Trichoplusia ni, respectively, but the isolate was essentially nontoxic to larvae of the beet armyworm, Spodoptera exigua. A bioassay of autoclaved spore-crystal preparations showed no evidence of ␤-exotoxin activity, indicating that toxicity was due primarily to Cry1Ab21. Studies of the pathogenesis of isolate IS5056 in second-instar larvae of T. ni showed that after larval death the bacterium colonized and subsequently sporulated extensively throughout the cadaver, suggesting that other bacteria inhabiting the midgut lumen played little if any role in mortality. As T. ni is among the most destructive pests of vegetable crops in North America and has developed resistance to B. thuringiensis, this new isolate may have applied value.

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Section: Discussionmentioning

confidence: 86%

Section: Methodsmentioning

confidence: 99%

Novel Isolate of Bacillus thuringiensis subsp. thuringiensis That Produces a Quasicuboidal Crystal of Cry1Ab21 Toxic to Larvae of Trichoplusia ni

Święcicka

Bideshi

Federici

2008

Appl Environ Microbiol

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“…The Cry2 and Cry3 proteins are 65 and 70 kDa, respectively. The dipteran active, Cry4 and Cry10 or Cry11 proteins are 135 and 80 kDa, respectively (Chambers et al 1991 Gough et al (2005) studied Bacillus thuringiensis strains collected from various Australian soil samples which produced crystals containing 130 and 28 kDa proteins. These strains were highly toxic to feeding larvae of sheep blowfly (Lucilia cuprina).…”

Section: Protein Profilingmentioning

confidence: 99%

Molecular characterization of indigenous Bacillus thuringiensis strains isolated from Kashmir valley

2017

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Bacillus thuringiensis (Bt) being an eco-friendly bioinsecticide is effectively used in pest management strategies and, therefore, isolation and identification of new strains effective against a broad range of target pests is important. In the present study, new indigenous B. thuringiensis strains were isolated and investigated so that these could be used as an alternative and/or support the current commercial strains/cry proteins in use. For this, 159 samples including soil, leaf and spider webs were collected from ten districts of Kashmir valley (India). Of 1447 bacterial strains screened, 68 Bt strains were identified with 4 types of crystalline inclusions. Crystal morphology ranking among the Bt strains was spherical (69.11%) [ spore attached (8.82%) [ rod (5.88%) = bipyramidal (5.88%) [ spherical plus rod (4.41%) [ spherical plus bipyramidal (2.94%) = cuboidal (2.94%). SDS-PAGE investigation of the spore-crystal mixture demonstrated Bt strains contained proteins of various molecular weights ranging from 150 to 28 kDa. Insecticidal activity of the 68 indigenous Bt strains against Spodoptera litura neonates showed that Bt strain SWK1 strain had the highest mortality. Lepidopteron active genes (cry1, cry2Ab, cry2Ab) were present in six Bt strains. Further, analysis of a full-length cry2A gene (*1.9 kb) by PCR-RFLP in strain SWK1 revealed that it was a new cry2A gene in Bt strain SWK1 and was named as cry2Al1 (GenBank Accession No. KJ149819.1) using the Bt toxin nomenclature (http://www.btnomenclature. info). Insect bioassays with neonate larvae of S. litura and H. armigera showed that the purified Cry2Al1 is toxic to S. litura with LC 50 2.448 lg/ml and H. armigera with LC 50 3.374 lg/ml, respectively. However, it did not produce any mortality in third instar larvae of Aedes aegypti, Culex quinquefasciatus and Anopheles stephensi larvae/pupae insects (100 lg/ml) at 28 ± 2°C and 75 to 85% relative humidity under a photoperiod of 14L:10D.

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“…However, sporulated cultures of these native microorganisms revealed protein bands of around 28 kDa. This molecular mass could correspond to Cyt toxins, entomocidal crystal proteins highly active against Diptera larvae (Gough et al 2005). On the other hand, all isolations with high toxic activity against Sf were located in cluster B.…”

Section: Discussionmentioning

confidence: 95%

Insecticidal crystal proteins from native Bacillus thuringiensis: numerical analysis and biological activity against Spodoptera frugiperda

et al. 2008

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Fourteen strains of Bacillus thuringiensis collected from both larvae showing disease symptoms and soil samples in northwest Argentina were characterized by insecticidal activity against Spodoptera frugiperda. First instar larvae and protein profile SDS-PAGE analysis of whole cell proteins not only allowed the differentiation of native Bacillus thuringiensis but also revealed the possibility of applying protein profile analysis in classification of toxicity patterns. Cluster analysis showed that there were two main groups. Interestingly, one of them only contained the most pathogenic native strains. The biomass-bound protease activity of native pathogenic isolates and the reference strain Bt 4D1 is also reported.

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Identification and characterization of proteins from Bacillus thuringiensis with high toxic activity against the sheep blowfly, Lucilia cuprina

Cited by 18 publications

References 25 publications

Novel Isolate of Bacillus thuringiensis subsp. thuringiensis That Produces a Quasicuboidal Crystal of Cry1Ab21 Toxic to Larvae of Trichoplusia ni

Novel Isolate of Bacillus thuringiensis subsp. thuringiensis That Produces a Quasicuboidal Crystal of Cry1Ab21 Toxic to Larvae of Trichoplusia ni

Molecular characterization of indigenous Bacillus thuringiensis strains isolated from Kashmir valley

Insecticidal crystal proteins from native Bacillus thuringiensis: numerical analysis and biological activity against Spodoptera frugiperda

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