Revision of the Nomenclature for the Bacillus thuringiensis Pesticidal Crystal Proteins (original) (raw)
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Microbiology and Molecular Biology Reviews
The crystal proteins of Bacillus thuringiensis have been extensively studied because of their pesticidal properties and their high natural levels of production. The increasingly rapid characterization of new crystal protein genes, triggered by an effort to discover proteins with new pesticidal properties, has resulted in a variety of sequences and activities that no longer fit the original nomenclature system proposed in 1989. Bacillus thuringiensis pesticidal crystal protein (Cry and Cyt) nomenclature was initially based on insecticidal activity for the primary ranking criterion. Many exceptions to this systematic arrangement have become apparent, however, making the nomenclature system inconsistent. Additionally, the original nomenclature, with four activity-based primary ranks for 13 genes, did not anticipate the current 73 holotype sequences that form many more than the original four subgroups. A new nomenclature, based on hierarchical clustering using amino acid sequence identi...
Bacillus thuringiensis and its pesticidal crystal proteins
Microbiology and molecular biology reviews : MMBR, 1998
During the past decade the pesticidal bacterium Bacillus thuringiensis has been the subject of intensive research. These efforts have yielded considerable data about the complex relationships between the structure, mechanism of action, and genetics of the organism's pesticidal crystal proteins, and a coherent picture of these relationships is beginning to emerge. Other studies have focused on the ecological role of the B. thuringiensis crystal proteins, their performance in agricultural and other natural settings, and the evolution of resistance mechanisms in target pests. Armed with this knowledge base and with the tools of modern biotechnology, researchers are now reporting promising results in engineering more-useful toxins and formulations, in creating transgenic plants that express pesticidal activity, and in constructing integrated management strategies to insure that these products are utilized with maximum efficiency and benefit.
Bacillus thuringiensis and Its Pesticidal Crystal Proteins
Microbiology and Molecular Biology Reviews, 1998
The leading biorational pesticide, Bacillus thuringiensis, is a ubiquitous gram-positive, spore-forming bacterium that forms a parasporal crystal during the stationary phase of its growth cycle. B. thuringiensis was initially characterized as an insect pathogen, and its insecticidal activity was attributed largely or completely (depending on the insect) to the parasporal crystals. This observation led to the development of bioinsecticides
Diversity of Insecticidal Crystal proteins (ICPs) of indigenous Bacillus thuringiensis strains
Tribhuvan University Journal of Microbiology, 2018
Objectives: The purpose of this study was to characterize the indigenous Bacillus thuringiensis (Bt) isolated from the soil samples of Terai. Methods: A total of 50 soil samples were collected from cultivated and barren fi elds of Terai region. Isolation was carried out using the acetate selection protocol Nutrient broth (NB) was acetated by using 0.25M sodium acetate which is a selective enrichment method for isolation of Bt. Characterization of the isolate was done by phenotyping methods (microscopy and biochemical). Results: No distinct variation was observed between the isolates of cultivable and uncultivable lands. Bt were categorized into 7 different types based on colony morphology. The dominant colony was fried egg type identical with the reference strain, followed by fl at white type of colony. The result showed that even though the colony morphology was same but the ICPs (Insecticidal crystal proteins) shapes produced by them varied, rod shapes (53.57%), spherical (10.71%), ovoid (8.3%), amorphous (17.85%), capheaded (9.5%). ICPs morphology revealed the cry1, cry2, cry3, cry4, cry8, cry 9, cry10 and cry11 types of gene may be present in the native isolates. Conclusion: This study represents the fi rst report of several indigenous Bacillus thuringiensis strains with signifi cantly different ICPs producing strains from hot tropical climate.
PCR-based identification of Bacillus thuringiensis pesticidal crystal genes
Fems Microbiology Reviews, 2003
The polymerase chain reaction (PCR) is a molecular tool widely used to characterize the insecticidal bacterium Bacillus thuringiensis. This technique can be used to amplify specific DNA fragments and thus to determine the presence or absence of a target gene. The identification of B. thuringiensis toxin genes by PCR can partially predict the insecticidal activity of a given strain. PCR has proven to be a rapid and reliable method and it has largely substituted bioassays in preliminary classification of B. thuringiensis collections. In this work, we compare the largest B. thuringiensis PCR-based screenings, and we review the natural occurrence of cry genes among native strains. We also discuss the use of PCR for the identification of novel cry genes, as well as the potential of novel technologies for the characterization of B. thuringiensis strains.
Applied and Environmental Microbiology, 2005
Bacillus thuringiensis crystal proteins of the Cry34 and Cry35 classes function as binary toxins showing activity on the western corn rootworm, Diabrotica virgifera virgifera LeConte. We surveyed 6,499 B. thuringiensis isolates by hybridization for sequences related to cry35A genes, identifying 78 strains. Proteins of the appropriate molecular mass (ca. 44 kDa) for Cry35 were observed in 42 of the strains. Full-length, or nearly full-length, sequences of 34 cry34 genes and 16 cry35 genes were also obtained from cloning, PCR analysis, and DNA sequencing. These included representatives of all known Cry34A, Cry34B, Cry35A, and Cry35B classes, as well as a novel Cry34A/Cry35A-like pair. Bioassay analysis indicated that cry35-hybridizing strains not producing a ca. 14-kDa protein, indicative of Cry34, were not active on corn rootworms, and that the previously identified Cry34A/Cry35A pairs were more active than the Cry34B/Cry35B pairs. The cry35-hybridizing B. thuringiensis strains were found in locales and materials typical for other B. thuringiensis strains. Comparison of the sequences with the geographic origins of the strains showed that identical, or nearly identical, sequences were found in strains from both Australasia and the Americas. Sequence similarity searches revealed that Cry34 proteins are similar to predicted proteins in Photorhabdus luminescens and Dictyostelium discoidium, and that Cry35Ab1 contains a segment similar to beta-trefoil domains that may be a binding motif. The binary Cry34/Cry35 B. thuringiensis crystal proteins thus appear closely related to each other, are environmentally ubiquitous, and share sequence similarities consistent with activity through membrane disruption in target organisms.
PCR analysis of the cryI insecticidal crystal family genes from Bacillus thuringiensis
Applied and environmental microbiology, 1994
A method allowing rapid and accurate identification of different subgroups within the insecticidal crystal CryI protein-producing family of Bacillus thuringiensis strains was established by using PCR technology. Thirteen highly homologous primers specific to regions within genes encoding seven different subgroups of B. thuringiensis CryI proteins were described. Differentiation among these strains was determined on the basis of the electrophoretic patterns of PCR products. B. thuringiensis strains, isolated from soil samples, were analyzed by PCR technology. Small amounts of bacterial lysates were assayed in two reaction mixtures containing six to eight primers. This method can be applied to rapidly detect the subgroups of CryI proteins that correspond with toxicity to various lepidopteran insects.
The novel δ-endotoxin from Bacillus thuringiensis (Bt) is one of the alternative measures for lepidopteran pest control. A survey of 31 Bt isolates, obtained from Thailand and previously screened for their toxicity to lepidopterans, was conducted to determine the presence of cry1-type genes, using polymerase chain reaction -restriction fragment length polymorphism analysis. Seven distinct types of cry1 genes: cry1Aa, cry1B, cry1C, cry1Cb, cry1D, cry1E, and cry1F, were identified. The most common of the cry1-type genes was cry1Cb, followed by cry1C and cry1D, which covered 64.6, 48.4, and 25.8%, respectively. Besides cry1C and cry1Cb, two candidate cry genes, cry1E and cry1D, of isolate JC 190 (harbouring cry1C/1E), with cotton bollworm toxin specificity, showed 99% amino acid sequence identity to Cry1Ea of B. thuringiensis subsp. kenyae, while isolate JC 291 (containing cry1C/1Cb/1D), with Asian corn borer toxin specificity, harboured Cry1Dc, which exhibited only 84% amino acid sequ...