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Molecular characterization of Bacillus thuringiensis using rep-PCR.

Rosane Bezerra da Silva1, Fernando Hercos Valicente2

  • 1Department of Plant Biotechnology, Federal University of Lavras, Lavras, Minas Gerais Brazil.

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|December 24, 2013
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Summary

Repetitive sequence-based PCR (Rep-PCR) revealed genetic divergence among 65 Bacillus thuringiensis (Bt) strains. The BOX primer was most informative, highlighting significant genetic variability within Bt populations.

Keywords:
Bacillus thuringiensisGenetic divergenceREP-PCRRepetitive sequences

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Bacillus thuringiensis (Bt) is a ubiquitous soil bacterium with significant agricultural and public health applications.
  • Understanding the genetic diversity of Bt strains is crucial for optimizing their use and developing new applications.
  • Repetitive sequence-based PCR (Rep-PCR) is a molecular technique used to assess genetic relatedness and diversity within bacterial populations.

Purpose of the Study:

  • To determine the genetic divergence among 65 Bacillus thuringiensis (Bt) strains using Repetitive sequence-based PCR (Rep-PCR).
  • To evaluate the informativeness of different primers (BOX, ERIC, REP) for differentiating Bt strains.
  • To analyze the population structure and genetic variability of Bt isolates from different sources.

Main Methods:

  • Sixty-five Bacillus thuringiensis (Bt) strains were analyzed using Repetitive sequence-based PCR (Rep-PCR).
  • Three primer sets (BOX, ERIC, and REP) were employed to amplify repetitive DNA sequences.
  • The generated DNA fragments were analyzed to construct a dendrogram for assessing genetic relatedness and population structure.

Main Results:

  • The BOX primer yielded the highest number of informative fragments (26), followed by ERIC (19) and REP (10), totaling 55 fragments.
  • A dendrogram revealed ten distinct genetic groups at an average distance of 45%, with Group 1 containing 41.5% of the isolates.
  • Bt strains from Embrapa predominantly clustered in Group 1 (53.2%), while strains from USDA and Institute Pasteur exhibited greater genetic variability.

Conclusions:

  • Rep-PCR, particularly with the BOX primer, is effective in delineating genetic diversity within Bacillus thuringiensis populations.
  • Significant genetic divergence exists among Bt strains, with notable differences observed between isolates from various collections.
  • The findings provide insights into the population structure of Bt, essential for strain selection and application development.