Differentiation and identification of iron-oxidizing acidophilic bacteria using cultivation techniques and amplified ribosomal DNA restriction enzyme analysis

D. Barrie Johnson, Naoko Okibe, Kevin B. Hallberg

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Acidophilic iron-oxidizing microorganisms are important both environmentally and in biotechnological applications. Although, as a group, they are readily detected by their ability to generate ferric iron (resulting in a distinctive color change in liquid media), these microbes highly diverse phylogenetically. Various other characteristics, such as optimum growth temperature, response to organic carbon sources, and cellular morphologies, facilitate, in some cases, identification of isolates to a genus or species level, although this approach has limitations and may give erroneous results. In this study, a combined approach of using physiological traits together with amplified ribosomal DNA restriction enzyme analysis (ARDREA) has been successful in identifying all known acidophilic iron-oxidizing bacteria to the species level. Computer-generated maps were used to identify restriction enzymes that allow the differentiation of the acidophiles, and these were confirmed experimentally using authentic bacterial strains. To test further the validity of this approach, six acidophilic moderately thermophilic iron-oxidizing bacteria isolated from Montserrat (West Indies) were analysed using the ARDREA protocol. Three of the isolates were identified as Sulfobacillus acidophilus-like, and one as Sulfobacillus thermosulfidooxidans-like bacteria. The fifth isolate gave DNA digest patterns that were distinct from all known strains of iron-oxidizing acidophiles. Subsequent sequencing of the 16S rRNA genes of these isolates confirmed the identity of the four Sulfobacillus isolates, and also that the fifth isolate was a novel species. Schematic diagrams showing how ARDREA may be used to rapidly identify all known acidophilic iron-oxidizing bacteria are presented.

Original languageEnglish
Pages (from-to)299-313
Number of pages15
JournalJournal of Microbiological Methods
Volume60
Issue number3
DOIs
Publication statusPublished - Jan 1 2005
Externally publishedYes

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Restriction Mapping
DNA Restriction Enzymes
Ribosomal DNA
Iron
Bacteria
West Indies
rRNA Genes
Carbon
Color
Temperature
DNA
Enzymes
Growth

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology
  • Microbiology (medical)

Cite this

Differentiation and identification of iron-oxidizing acidophilic bacteria using cultivation techniques and amplified ribosomal DNA restriction enzyme analysis. / Johnson, D. Barrie; Okibe, Naoko; Hallberg, Kevin B.

In: Journal of Microbiological Methods, Vol. 60, No. 3, 01.01.2005, p. 299-313.

Research output: Contribution to journalArticle

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