Characterization of Bacillus thuringiensis and related bacteria by ribosomal RNA gene restriction fragment length polymorphisms

Priest, Fergus G.; Kaji, Denise A.; Rosato, Yoko Bomura; Canhos, Vanderlei Perez

doi:10.1099/13500872-140-5-1015

Cited by 77 publications

(52 citation statements)

References 31 publications

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“…Some important B. thuringiensis subspecies known to be genetically related were grouped together by ITS-HHP. For example, strains belonging to the B. thuringiensis subspecies aizawai, galleriae, kurstaki, and tolworthi showed the same ITS-HHP patterns, in agreement with the ribotyping data of Priest et al (49), the M13 DNA fingerprinting data of Miteva et al (40), and the DNA-DNA hybridization data of Nakamura (42). The fact that the ITS-HPP patterns of these subspecies were identical reflected their close DNA relatedness (90%) (42), while the B. thuringiensis subspecies darmstadiensis, israelensis, and sotto, which have relatively low DNA-DNA hybridization levels (below 70%) (42), showed different ITS-HPP types and were assigned to a cluster different from that of B. thuringiensis subsp.…”

supporting

confidence: 78%

“…Agarose gel ITS-PCR fingerprinting patterns are identical for the six species of the B. cereus group (14, 17) and are thus not suitable for the rapid identification of the species. Since the B. cereus group strains have from 8 to 12 ribosomal operons (34,39,49) that can differ in ITS sequence, it might be possible to detect interspecies and intraspecies differences through ITS homoduplex-heteroduplex polymorphisms (ITS-HHP) (6, 31-33). The principle of strain discrimination by ITS-HHP analysis is based on the detection of sequence polymorphisms in the form of heteroduplex DNA bands that are generated during PCR when fragments of different length, which contain homologous regions at the 3Ј and 5Ј ends, cross-hybridize to form heteroduplex DNA structures.…”

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confidence: 99%

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Homoduplex and Heteroduplex Polymorphisms of the Amplified Ribosomal 16S-23S Internal Transcribed Spacers Describe Genetic Relationships in the “ Bacillus cereus Group”

Daffonchio

Chérif

Borin

2000

Appl Environ Microbiol

142

155

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Bacillus anthracis, Bacillus cereus, Bacillus mycoides, Bacillus pseudomycoides, Bacillus thuringiensis, and Bacillus weihenstephanensis are closely related in phenotype and genotype, and their genetic relationship is still open to debate. The present work uses amplified 16S-23S internal transcribed spacers (ITS) to discriminate between the strains and species and to describe the genetic relationships within the "B. cereus group," advantage being taken of homoduplex-heteroduplex polymorphisms (HHP) resolved by polyacrylamide gel electrophoresis and silver staining. One hundred forty-one strains belonging to the six species were investigated, and 73 ITS-HHP pattern types were distinguished by MDE, a polyacrylamide matrix specifically designed to resolve heteroduplex and single-strand conformation polymorphisms. The discriminating bands were confirmed as ITS by Southern hybridization, and the homoduplex or heteroduplex nature was identified by single-stranded DNA mung bean nuclease digestion. Several of the ITS-HHP types corresponded to specific phenotypes such as B. anthracis or serotypes of B. thuringiensis. Unweighted pair group method arithmetic average cluster analysis revealed two main groups. One included B. mycoides, B. weihenstephanensis, and B. pseudomycoides. The second included B. cereus and B. thuringiensis, B. anthracis appeared as a lineage of B. cereus.The "Bacillus cereus group," one of the most homogeneous groups of the genus Bacillus, encompasses six validly described species, B. anthracis, B. cereus, B. mycoides, B. pseudomycoides, B. thuringiensis, and B. weihenstephanensis, all of which have an important impact on human activity (18,39,44).These species are known to be strictly related phylogenetically, as has been shown by DNA-DNA hybridization studies (35,39,(42)(43)(44)57) and the sequencing of the ribosomal RNA genes (2-4, 39). On the other hand, a marked variability is always observed when large collections of strains are examined by DNA fingerprinting methods that target the whole genome (8-13, 17, 25-29, 36, 58) and/or discrete genes (15,16,38,54,55,61). Hence, the phylogenetic and taxonomic relationship among these species is still open to debate. It has been proposed previously that B. anthracis, B. cereus, and B. thuringiensis represent a single species, this conclusion having been reached through genome sizing and mapping (9-13) and, very recently, by multilocus enzyme electrophoresis and the sequencing of nine different DNA loci (27).The ribosomal operon is a classic molecular marker used to trace genetic relationships and to identify strains rapidly (1). Of all the different regions of the ribosomal operon, the internal transcribed spacers (ITS) between 16S and 23S ribosomal DNA are frequently used as molecular markers to identify microbial species and analyze the phylogenetic relationship between strains (see, for example, references 14, 22, and 24). ITS are generally found in multiple copies in most bacterial genomes (24, 33). Since ITS are hypervariable with respect to adjace...

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supporting

confidence: 78%

mentioning

confidence: 99%

Homoduplex and Heteroduplex Polymorphisms of the Amplified Ribosomal 16S-23S Internal Transcribed Spacers Describe Genetic Relationships in the “ Bacillus cereus Group”

Daffonchio

Chérif

Borin

2000

Appl Environ Microbiol

142

155

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“…Some strains produce extracellular enterotoxins and cause food poisoning in man, and other toxins such as phospholipase C, haemolysins and emetic toxins can also be produced. B. thuringiensis is characterized by the synthesis of highly specific insecticidal toxins and is one of the most widely used biopesticides (Priest et al, 1994 organism and is identified unambiguously by its unique rhizoid colony morphology and its lack of motility. Finally, B. antbracis is the causative agent of anthrax in humans and animals.…”

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confidence: 99%

Diversity and differential distribution of IS231, IS232 and IS240 among Bacillus cereus, Bacillus thuringiensis and Bacillus mycoides

1997

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Bacillus cereus, Bacillus thuringiensis and Bacillus mycoides are very closely related bacteria, generally considered as subspecies of B. cereus sensu lato. Different transposable elements have been isolated from B. thuringiensis, including IS231, IS232 and IS240 and their variants. The distribution of these three insertion sequences (IS) within the B. cereus group has been investigated in 90 strains of B. thuringiensis (representing 61 serovars), in 30 reference strains of B. cereus and in 33 strains of B. mycoides. Since these IS elements are delimited by well-conserved and specific inverted repeats, the use of primers corresponding to these ends allowed their amplification by PCR. The results showed that IS231 is the most abundant element in the three taxa, whereas IS232 is apparently exclusively associated with B. thuringiensis. Hybridization and Dral RFLP analysis of the PCR products confirmed and extended knowledge of the heterogeneity previously observed among iso-IS231 elements. Moreover, a similar diversity was observed among iso-IS240 elements. This contrasted with the relative homogeneity displayed by iso-IS232 elements. No specific association appeared to exist between any particular iso-element and a specific strain or serotype.

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“…We identified four classes of PlcR-PapR pairs, defining four distinct pherotypes in the B. cereus group. We used these findings to look at the evolution of the PlcR-PapR quorum-sensing system with regard to the phylogeny of the species forming the B. cereus group.The Bacillus cereus group includes six highly related species of gram-positive, spore-forming, AT-rich bacteria: Bacillus anthracis, Bacillus cereus (sensu stricto), Bacillus mycoides, Bacillus pseudomycoides, Bacillus thuringiensis, and Bacillus weihenstephanensis (15,18,21). These species were originally identified on the basis of phenotypic traits, such as the production of a capsule and toxins (B. anthracis), the rhyzoid morphology of the colonies (B. mycoides and B. pseudomycoides),…”

mentioning

confidence: 99%

“…The Bacillus cereus group includes six highly related species of gram-positive, spore-forming, AT-rich bacteria: Bacillus anthracis, Bacillus cereus (sensu stricto), Bacillus mycoides, Bacillus pseudomycoides, Bacillus thuringiensis, and Bacillus weihenstephanensis (15,18,21). These species were originally identified on the basis of phenotypic traits, such as the production of a capsule and toxins (B. anthracis), the rhyzoid morphology of the colonies (B. mycoides and B. pseudomycoides), the production of parasporal inclusions (B. thuringiensis), and psychrotolerance (B. weihenstephanensis).…”

mentioning

confidence: 99%

Specificity and Polymorphism of the PlcR-PapR Quorum-Sensing System in theBacillus cereusGroup

Slamti

Lereclus

2005

J Bacteriol

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The expression of extracellular virulence factors in various species of the Bacillus cereus group is controlled by the plcR and papR genes, which encode a transcriptional regulator and a cell-cell signaling peptide, respectively. A processed form of PapR, presumably a pentapeptide, specifically interacts with PlcR to facilitate its binding to its DNA targets. This activating mechanism is strain specific, with this specificity being determined by the first residue of the pentapeptide. We carried out in vivo complementation assays and compared the PlcRPapR sequences of 29 strains from the B. cereus group. Our findings suggested that the fifth amino acid of the pentapeptide is also involved in the specificity of activation. We identified four classes of PlcR-PapR pairs, defining four distinct pherotypes in the B. cereus group. We used these findings to look at the evolution of the PlcR-PapR quorum-sensing system with regard to the phylogeny of the species forming the B. cereus group.The Bacillus cereus group includes six highly related species of gram-positive, spore-forming, AT-rich bacteria: Bacillus anthracis, Bacillus cereus (sensu stricto), Bacillus mycoides, Bacillus pseudomycoides, Bacillus thuringiensis, and Bacillus weihenstephanensis (15,18,21). These species were originally identified on the basis of phenotypic traits, such as the production of a capsule and toxins (B. anthracis), the rhyzoid morphology of the colonies (B. mycoides and B. pseudomycoides),

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Characterization of Bacillus thuringiensis and related bacteria by ribosomal RNA gene restriction fragment length polymorphisms

Cited by 77 publications

References 31 publications

Homoduplex and Heteroduplex Polymorphisms of the Amplified Ribosomal 16S-23S Internal Transcribed Spacers Describe Genetic Relationships in the “ Bacillus cereus Group”

Homoduplex and Heteroduplex Polymorphisms of the Amplified Ribosomal 16S-23S Internal Transcribed Spacers Describe Genetic Relationships in the “ Bacillus cereus Group”

Diversity and differential distribution of IS231, IS232 and IS240 among Bacillus cereus, Bacillus thuringiensis and Bacillus mycoides

Specificity and Polymorphism of the PlcR-PapR Quorum-Sensing System in theBacillus cereusGroup

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