Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Application of Molecular Methods for the Identification of Acetic Acid Bacteria Isolated from Blueberries and Citrus Fruits

  • Gerard, Liliana Mabel (Laboratorio de Microbiologia y Biotecnologia de Alimentos, Facultad de Ciencias de la Alimentacion, Universidad Nacional de Entre Rios) ;
  • Davies, Cristina Veronica (Laboratorio de Microbiologia y Biotecnologia de Alimentos, Facultad de Ciencias de la Alimentacion, Universidad Nacional de Entre Rios) ;
  • Solda, Carina Alejandra (Laboratorio de Microbiologia y Biotecnologia de Alimentos, Facultad de Ciencias de la Alimentacion, Universidad Nacional de Entre Rios) ;
  • Corrado, Maria Belen (Laboratorio de Microbiologia y Biotecnologia de Alimentos, Facultad de Ciencias de la Alimentacion, Universidad Nacional de Entre Rios) ;
  • Fernandez, Maria Veronica (Laboratorio de Microbiologia y Biotecnologia de Alimentos, Facultad de Ciencias de la Alimentacion, Universidad Nacional de Entre Rios)
  • Received : 2019.12.14
  • Accepted : 2020.01.23
  • Published : 2020.06.28
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Abstract

Sixteen acetic acid bacteria (AAB) were isolated from blueberries and citric fruits of the Salto Grande region (Concordia, Entre Rios, Argentina) using enrichment techniques and plate isolation. Enrichment broths containing ethanol and acetic acid enabled maximum AAB recovery, since these components promote their growth. Biochemical tests allowed classification of the bacteria at genus level. PCR-RFLP of the 16S rRNA and PCR-RFLP of the 16S-23S rRNA intergenic spacer allowed further classification at the species level; this required treatment of the amplified products of 16S and 16S-23S ITS ribosomal genes with the following restriction enzymes: AluI, RsaI, HaeIII, MspI, TaqI, CfoI, and Tru9I. C7, C8, A80, A160, and A180 isolates were identified as Gluconobacter frateurii; C1, C2, C3, C4, C5, C6, A70, and A210 isolates as Acetobacter pasteurianus; A50 and A140 isolates as Acetobacter tropicalis; and C9 isolate as Acetobacter syzygii. The bacteria identified by 16S rRNA PCR-RFLP were validated by 16S-23S PCR-RFLP; however, the C1 isolate showed different restriction patterns during identification and validation. Partial sequencing of the 16S gene resolved the discrepancy.

Keywords

References

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