Probing the archaeal diversity of a mixed thermophilic bioleaching culture by TGGE and FISH

Mikkelsen, Deirdre, Kappler, Ulrike, McEwan, Alastair G. and Sly, Lindsay I. (2009) Probing the archaeal diversity of a mixed thermophilic bioleaching culture by TGGE and FISH. Systematic and Applied Microbiology, 32 7: 501-513. doi:10.1016/j.syapm.2009.06.001

Author Mikkelsen, Deirdre
Kappler, Ulrike
McEwan, Alastair G.
Sly, Lindsay I.
Title Probing the archaeal diversity of a mixed thermophilic bioleaching culture by TGGE and FISH
Journal name Systematic and Applied Microbiology   Check publisher's open access policy
ISSN 0723-2020
Publication date 2009-10
Sub-type Article (original research)
DOI 10.1016/j.syapm.2009.06.001
Volume 32
Issue 7
Start page 501
End page 513
Total pages 13
Editor Thomas Bosch
Place of publication Jena, Germany
Publisher Urban and Fischer Verlag
Collection year 2010
Language eng
Subject C1
060501 Bacteriology
060504 Microbial Ecology
840205 Mining and Extraction of Precious (Noble) Metal Ores
849899 Environmentally Sustainable Mineral Resource Activities not elsewhere classified
0603 Evolutionary Biology
Formatted abstract
The archaeal community present in a sample of Mixed Thermophilic Culture-B (MTC-B) from a laboratory-scale thermophilic bioleaching reactor was investigated by temperature gradient gel electrophoresis (TGGE) and fluorescence in situ hybridisation (FISH). Both techniques were specifically adapted for use on native state bioleaching samples, with a view to establishing convenient means for monitoring culture composition. Using the TGGE protocol developed, the relative species composition of the thermophilic bioleaching sample was analysed, and included four phylotypes belonging to the Sulfolobales, which were related to Stygiolobus azoricus, Metallosphaera sp. J1, Acidianus infernus and Sulfurisphaera ohwakuensis. However, the St. azoricus-like phylotype was difficult to resolve and some micro-heterogeneity was observed within this phylotype. Specific FISH probes were designed to qualitatively assess the presence of the phylotypes in MTC-B. The sample was dominated by Sf. ohwakuensis-like Archaea. In addition, the St. azoricus-like, Metallosphaera species-like and Acidianus species-like cells appeared in similar low abundance in the community. Most strikingly, FISH identified Sulfolobus shibatae-like cells present in low numbers in the sample even though these were not detected by PCR-dependent TGGE. These results highlight the importance of using more than one molecular technique when investigating the archaeal diversity of complex bioleaching reactor samples.
© 2009 Elsevier GmbH. All rights reserved.
Keyword Bioleaching
Fluorescence in situ hybridisation
Molecular ecology
Temperature gradient gel electrophoresis
Q-Index Code C1
Q-Index Status Confirmed Code
Additional Notes Author prepress title: "Analysis of archaeal diversity in a mixed thermophilic bioleaching culture by TGGE and FISH".

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Created: Thu, 12 Nov 2009, 11:55:56 EST by Mr Andrew Martlew on behalf of School of Chemistry & Molecular Biosciences