A phylogenetic analysis of Bacillus thuringiensis serovars by RFLP-based ribotyping

Joung, K.‐B.; Côté, Jean-Charles

doi:10.1046/j.1365-2672.2001.01388.x

Cited by 12 publications

(7 citation statements)

References 35 publications

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“…Diversity of rRNA intergenic spacer sequences of 31 strains proved insufficient to discriminate between isolates (97 to 99% similarity) (6). On the other hand, ribotyping (16S, 23S, and 5S rRNA gene RFLP) of 80 serovars of B. thuringiensis showed a great diversity of patterns (27,28), similar to the diversity found with fluorescent AFLP, when 34 B. thuringiensis serovars were analyzed along with strains of B. cereus and B. anthracis (21).…”

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confidence: 79%

Fingerprinting of Bacillus thuringiensis Type Strains and Isolates by Using Bacillus cereus Group-Specific Repetitive Extragenic Palindromic Sequence-Based PCR Analysis

Reyes-Ramírez

Ibarra

2005

Appl Environ Microbiol

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A total of 119 Bacillus thuringiensis strains (83 type strains and 26 native isolates), as well as five B. cereus group species, were analyzed by repetitive extragenic palindromic sequence-based PCR analysis (Rep-PCR) fingerprinting. Primers Bc-REP-1 and Bc-REP-2 were specifically designed according to an extragenic 26-bp repeated sequence found in the six B. cereus group genomes reported. A total of 47 polymorphic bands were detected, and the patterns varied from 5 to 13 bands in number and from 0.2 to 3.8 kb in size. Virtually each type strain showed a distinctive B. cereus (Bc)-Rep-PCR pattern, except for B. thuringiensis serovars dakota (H serotype 15 [H15]) and sotto (H4a,4b), as well as serovars amagiensis (H29) and seoulensis (H35), which shared the same patterns. As expected, serovar entomocidus (H6) and its biovar subtoxicus showed an identical pattern; similarly, serovars sumiyoshiensis (H3a,3d) and fukuokaensis (H3a,3d,3e), which share two antigenic determinants, also showed identical Bc-Rep-PCR patterns. Interestingly, serovars israelensis (H14) and malaysiensis (H36), which share several phenotypic attributes, also showed identical Bc-Rep-PCR patterns. Native, coleopteran-active strains, including the self-agglutinated LBIT-74 strain, showed Bc-Rep-PCR patterns identical or very similar to that of the tenebrionis strain. Likewise, native mosquitocidal strains (including some self-agglutinated strains) also showed patterns identical or very similar to that of the serovar israelensis IPS-82 strain. Additionally, native ␤-exotoxin-producing strains from serovar thuringiensis showed patterns identical to that of the B. thuringiensis type strain. The B. cereus group-specific Bc-Rep-PCR fingerprinting technique was shown to be highly discriminative, fast, easy, and able to identify B. thuringiensis serotypes, including nonflagellar and self-agglutinated strains.

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confidence: 79%

Fingerprinting of Bacillus thuringiensis Type Strains and Isolates by Using Bacillus cereus Group-Specific Repetitive Extragenic Palindromic Sequence-Based PCR Analysis

Reyes-Ramírez

Ibarra

2005

Appl Environ Microbiol

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“…All three strains in cluster C belonged to group III. Therefore, there were some similarities and some differences between our GP results and those of Joung and Côté (2001b).…”

Section: Genome Profiling Analysismentioning

confidence: 50%

“…The phylogenetic relationships determined using GP were compared with those obtained from the RFLP analysis of rRNA gene sequences conducted by Joung and Côté (2001b) (Fig. 3).…”

Section: Genome Profiling Analysismentioning

confidence: 99%

Phylogenetic relationships among Bacillus thuringiensis (Bacillaceae: Bacillales) strains based on a comparison of SSU rRNA sequences and genome profiling

2011

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Bacillus thuringiensis Berliner has previously been classified via the serological identification of flagellar antigens. However, the phylogenetic relationships among strains of B. thuringiensis cannot be investigated by serotyping. Furthermore, high levels of homology have been found in gene sequences among various strains, complicating the determination of their evolutionary relationships. In order to elucidate the phylogenetic relationships within B. thuringiensis, we analyzed 40 strains belonging to typical serotypes using two approaches: an analysis of small subunit (SSU) rRNA sequences and genome profiling (GP) based on temperature gradient gel electrophoresis of random PCR products. The SSU rRNA analysis resulted in all 40 strains forming a single cluster with Bacillus cereus Frankland & Frankland. The distances among the subclusters were too small to further classify the strains. On the other hand, the phylogenetic analysis based on GP resulted in three clusters of B. thuringiensis strains. These results suggest that GP is a better method for the determination of phylogenetic relationships within B. thuringiensis.

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“…The high degree of internal divergence inside the minisatellite suggests that this is a very old tandem repeat and that no insertion or deletion events occurred in the recent past. This high internal heterogeneity is likely to further prevent the production of new al- (21). In Fig.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Minisatellite Polymorphisms in the Bacillus cereus Complex: a Simple Assay for Large-Scale Screening and Identification of Strains Most Closely Related to Bacillus anthracis

Valjevac

Hilaire

Lisanti

et al. 2005

Appl Environ Microbiol

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Polymorphism of five tandem repeats that are monomorphic in Bacillus anthracis was investigated in 230 isolates of the B. cereus group and in 5 sequenced B. cereus genomes in search for markers allowing identification of B. cereus and B. thuringiensis strains most closely related to B. anthracis. Using this multiple-locus variable number of tandem repeat analysis (MLVA), a cluster of 30 strains was selected for further characterization. Eventually, six of these were characterized by multilocus sequence type analysis. One of the strains is only six point mutations (of almost 3,000 bp) away from B. anthracis and was also proposed to be closest to B. anthracis by MLVA analysis. However, this strain remains separated from B. anthracis by a number of significant genetic events observed in B. anthracis, including the loss of the hemolysin activity, the presence of four prophages, and the presence of the two virulence plasmids, pXO1 and pXO2. One particular minisatellite marker provides an efficient assay to identify the subset of B. cereus and B. thuringiensis strains closely related to B. anthracis. Based on these results, a very simple assay is proposed that allows the screening of hundreds of strains from the B. cereus complex, with modest equipment and at a low cost, to eventually fill the gap with B. anthracis and better understand the origin and making of this dangerous pathogen.

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A phylogenetic analysis of Bacillus thuringiensis serovars by RFLP-based ribotyping

Cited by 12 publications

References 35 publications

Fingerprinting of Bacillus thuringiensis Type Strains and Isolates by Using Bacillus cereus Group-Specific Repetitive Extragenic Palindromic Sequence-Based PCR Analysis

Fingerprinting of Bacillus thuringiensis Type Strains and Isolates by Using Bacillus cereus Group-Specific Repetitive Extragenic Palindromic Sequence-Based PCR Analysis

Phylogenetic relationships among Bacillus thuringiensis (Bacillaceae: Bacillales) strains based on a comparison of SSU rRNA sequences and genome profiling

Comparison of Minisatellite Polymorphisms in the Bacillus cereus Complex: a Simple Assay for Large-Scale Screening and Identification of Strains Most Closely Related to Bacillus anthracis

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