Influence of Ephestia kuehniella stage larvae on the potency of Bacillus thuringiensis Cry1Aa delta-endotoxin (original) (raw)
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Molecular biotechnology, 2008
A comparative study of different steps in the mode of action of the individual Bacillus thuringiensis kurstaki BNS3 Cry1Aa and Cry1Ac d-endotoxins on E. kuehniella larvae was performed in order to investigate the origin of the difference in the response of this larvae to each of the latter. Proteolytic activation was shown to be one of the main steps impaired in E. kuehniella tolerance to Cry1Aa. The absence of two proteinase activities as well as an altered activity level observed in the case of Cry1Aa would be the consequence of proteinase-mediated tolerance of E. kuehniella to this toxin. In situ binding and histopathological effect analyses allowed concluding that the binding of the toxin to BBMV receptors is the key step in E. kuehniella tolerance to Cry1Aa toxin. The latter was slightly bound to apical membranes of epithelial cells that remained intact, whereas Cry1Ac was tightly bound to completely damaged cells basal membranes.
Biological Control, 2006
Photorhabdus temperata strain K122 exhibited oral toxicity against Prays oleae with an LC 50 of 58.1 £ 10 6 cells ml ¡1 . Recombinant P. temperata strains expressing the cry1Aa and/or cry1Ia genes of Bacillus thuringiensis have been constructed. The two cry genes, encoding -endotoxins, were placed under the control of the lac promoter and IPTG dependent expression in P. temperata was demonstrated. The presence of the cry genes in K122 resulted in a clear improvement of oral toxicity. This improvement was of 6.2-, 6.6-, and 14.6-fold for the strains K122(pBCcry1Aa), K122(pBScry1Ia), and K122(pBCcry1Aa + pBScry1Ia), respectively. Furthermore, determination of the Synergistic Factor between Cry1Aa and Cry1Ia showed that they act synergistically. This work demonstrates that the heterologous expression of B. thuringiensis cry genes in P. temperata can be used to improve and broaden its host range for insect control.
World Journal Of Advanced Research and Reviews, 2021
Insect pests represent a major threat to food crops and human health, and therefore have to be combated in several ways, including chemical methods. However, researchers demonstrated that these molecules are dangerous for the farmers, consumers and the environment in general. For this reason, scientists permanently searched environment friendly alternatives such as the use of the bacterium Bacillus thuringiensis classified as one of the best insect pathogens. This microorganism is known by its ability to produce two types of insecticidal proteins, Vegetative insecticidal proteins (Vip) and delta-endotoxins produced during vegetative and sporulation stages of growth, respectively. In the present study, 15 B. thuringiensis strains were isolated from soil collected from different regions in Saudi Arabia (Al Baha, Jeddah, Khulis and Yanbu). B. thuringiensis isolates were then classified according to the shape of their parasporal crystals identified under microscope and proteins content of these crystals. Delta-endotoxins efficiency of the different isolates was investigated and promising strains were identified as very active. After 5 days-treatment, B. thuringiensis isolates 14 and 7 killed Ephestia kuehniella larvae with low LC50 of about 59.18 and 65.67 g/cm 2 , respectively. The results described in the present study proved that the new B. thuringiensis isolates could be of a great interest in the control of lepidopteran pests by using their delta-endotoxins in bioinsecticide formulations.
Biological Control, 2005
To determine the optimal ratio of Bacillus thuringiensis delta-endotoxins Cry1Aa, Cry1Ac, and Cry2Aa bioinsecticide formulations, and for development of novel biopesticides based on B. thuringiensis, the toxicities of these three proteins, individually and in combinations, have been determined against the Mediterranean Xour moth, Ephestia kuehniella. While B. thuringiensis crystals containing a mixture of Cry1Aa, Cry1Ac, and Cry2Aa displayed toxicity with an LC 50 and LC 95 of 109.7 and 463.0 ng of toxin per mg Xour, respectively, when used individually or in combination, Cry1Aa, Cry1Ac, and Cry2Aa showed signiWcantly lower activity. Thus, it is clear that synthesis of these toxins together in the wild strain is more eYcient than a mixture of them.
Archives of Microbiology, 2014
Insecticides derived from Bacillus thuringiensis are gaining worldwide importance as environmentally desirable alternatives to chemicals for the control of pests in public health and agriculture. Isolation and characterization of new strains with higher and broader spectrum of activity is an ever growing field. In the present work, a novel Tunisian B. thuringiensis isolate named BLB459 was characterized and electrophoresis assay showed that among a collection of 200 B. thuringiensis strains, the plasmid profile of BLB459 was distinctive. SmaI-PFGE typing confirmed the uniqueness of the DNA pattern of this strain, compared with BUPM95 and HD1 reference strains. PCR and sequencing assays revealed that BLB459 harbored three cry genes (cry30, cry40 and cry54) corresponding to the obtained molecular sizes in the protein pattern. Interestingly, PCR-RFLP assay demonstrated the originality of the BLB459 cry30-type gene compared to the other published cry30 genes. Insecticidal bioassays showed that BLB459 spore-crystal suspension was highly toxic to both Ephestia kuehniella and Spodoptera littoralis with LC50 values of about 64 (53-75) and 80 (69-91) µg of toxin cm-2 , respectively, comparing with that of the commercial strain HD1 used as reference. Important histopathological effects of BLB459 δ-endotoxins on the two tested larvae midguts were detected, traduced by the vacuolization of the apical cells, the damage of microvilli, and the disruption of epithelial cells. These results proved that BLB459 strain could be of a great interest for lepidopteran biocontrol.
A total of 73 Bacillus thuringiensis (Bt) strains were screened for the presence of non-hemolytic insecticidal -exotoxin-free -endotoxins. Out of them, 45 Bt strains produced -endotoxins with specific insecticidal activity against Drosophila melanogaster and/or Ephestia kuhniella larvae. The thermostable -exotoxin was observed only in 15 Bt strains and appeared to exhibit dual non-specific insecticidal activity against both D. melanogaster and E. kuhniella larvae and showed in vitro hemolysis for human erythrocytes. It was found that -exotoxin was produced by Bt strains belonging to five serovars (israelensis, kenyae, kurstaki, pakistani, and tohokuensis) and two non-serotypable strains. This result suggests that β-exotoxin production is a strain-specific property rather than a serovar-specific property. To our knowledge, this is the first study that demonstrates -exotoxins production association with Bt strains belonging to serovars israelensis, pakistani, and tohokuensis. Th...
Biochemical and biophysical research communications, 2017
The basis of the different susceptibility of Ephestia kuehniella to the Cry1Aa and Cry1Ac δ-endotoxins from Bacillus thuringiensis kurstaki BNS3 was studied. Both toxins bound specifically to the BBMV of E. kuehniella. The result of the ligand blot showed that Cry1Ac bound to three putative receptors of about 100, 65 and 80 kDa and Cry1Aa interacted only with a 100 kDa protein. Pronase digestion of the BBMV-bound toxins was used to analyze the toxin insertion. Both toxins inserted into the BBMV as monomers however, a 14 kDa peptide of α4-α5 which correspond to the oligomeric form of this peptide was detected in case of Cry1Ac only. Analysis of the in vitro oligomerisation of these toxins in the presence of the BBMV of E. kuehniella showed reduced oligomer formation in case of Cry1Aa in comparison with Cry1Ac. Taken together, these results strongly suggest that the difference of toxicity between Cry1Aa and Cry1Ac to E. kuehniella is due to a deficient oligomerisation of Cry1Aa.
The study of 257 crystal-producing Bacillus thuringiensis isolates from bioinsecticide free soil samples collected from different sites in Tunisia, was performed by PCR amplification, using six primer pairs specific for cry1, cry2, cry3, cry4, and vip3A genes, by the investigation of strain plasmid pattern, crystal morphology and deltaendotoxin content and by the assessment of insecticidal activities against the lepidopteran insect Ephestia kuehniella. Based on plasmid pattern study, 11 representative strains of the different classes were subjected to morphological and molecular analyses. The comparison of the PFGE fingerprints confirmed the heterogeneity of these strains. B. thuringiensis kurstaki strains, harbouring at the same time the genes cry1A, cry2, cry1Ia, and vip3A, were the most abundant (65.4%). 33.34% of the new isolates showed particular delta-endotoxin profiles but no PCR products with the used primer sets. B. thuringiensis israelensis was shown to be also very rare among the Tunisian B. thuringiensis isolates diversity. These findings could have considerable impacts for the set up of new pest control biological agents.
African Journal of Biotechnology, 2005
Bacillus thuringiensis was investigated in four different habitats (grain dust, olive-cultivated soils, waste and industrial-byproducts contaminated soils, and animal byproducts-contaminated soils). The bacterium was highly abundant in soils contaminated with animal byproducts. Eight serotypes with Bacillus thuringiensis israelensis being the most common. Out of the twenty-six isolated strains, five strains (serotype: kenyae, kurstaki, kurstaki HD1 and thuringiensis) that produced bipyramid crystal proteins were toxic to the lepidoptera larvae of Ephestia kuehniella Zeller. The SDS-PAGE protein profile analysis showed a relationship between the crystal protein shape and the toxicity to the larvae of the tested insect.
AFRICAN JOURNAL OF BIOTECHNOLOGY, 2012
A total of 73 Bacillus thuringiensis (Bt) strains were screened for the presence of non-hemolytic insecticidal -exotoxin-free -endotoxins. Out of them, 45 Bt strains produced -endotoxins with specific insecticidal activity against Drosophila melanogaster and/or Ephestia kuhniella larvae. The thermostable -exotoxin was observed only in 15 Bt strains and appeared to exhibit dual non-specific insecticidal activity against both D. melanogaster and E. kuhniella larvae and showed in vitro hemolysis for human erythrocytes. It was found that -exotoxin was produced by Bt strains belonging to five serovars (israelensis, kenyae, kurstaki, pakistani, and tohokuensis) and two non-serotypable strains. This result suggests that β-exotoxin production is a strain-specific property rather than a serovarspecific property. To our knowledge, this is the first study that demonstrates -exotoxins production association with Bt strains belonging to serovars israelensis, pakistani, and tohokuensis. The plasmid DNA profiles of some -exotoxin producing Bt strains shared large plasmid patterns which may have the common -exotoxin regulatory gene(s). It was found that 16 local Bt strains, 15 of which belonged to five serovars (aizawai, israelensis, kurstaki, morrisoni, and pakistani) and one was autoagglutinated strain, produced non-hemolytic insecticidal -exotoxin-free -endotoxins. Based on random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), the genotypic relatedness among these 16 Bt strains was investigated. The strains were grouped into two clusters. Bt strains within serovars israelensis were grouped in two subclusters, Bt strains within serovars aizawai were genomically homogeneous and clustered together, while the other serovars were grouped together in one subcluster. The autoagglutinated strain was clustered within serovar israelensis. Thus, these endotoxins can be developed for the use in Bt-based insecticidal preparations.