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Ribosomal intergenic spacer analysis as a tool for monitoring methanogenic archaea changes in an anaerobic digester

Abstract

The applicability of a newly-designed PCR primer pair in examination of methanogenic Archaea in a digester treating plant biomass was evaluated by Ribosmal Intergenic Spacer Analysis (RISA). To find a suitable approach, three variants of RISA were tested: (1) standard, polyacrylamide gel-based, (2) automated, utilized capillary electrophoresis (GA-ARISA), and (3) automated microfluidics-based (MF-ARISA). All three techniques yielded a consistent picture of archaeal community structure changes during anaerobic digestion monitored for more than 6 weeks. While automated variants were more practical for handling and rapid analysis of methanogenic Archaea, the gel-based technique was advantageous when micro-organism identification was required. A DNA-sequence analysis of dominant bands extracted from the gel revealed that the main role in methane synthesis was played by micro-organisms affiliated with Methanosarcina barkeri. The obtained results revealed that RISA is a robust method allowing for detailed analysis of archaeal community structure during organic biomass conversion into biogas. In addition, our results showed that GA-ARISA has a higher resolution and reproducibility than other variants of RISA and could be used as a technique for tracking changes in methanogenic Archaea in an anaerobic digester.

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DOI:
Digital Object Identifier (open in new tab) 10.1007/s00284-013-0353-2
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Category:
Articles
Type:
artykuł w czasopiśmie wyróżnionym w JCR
Published in:
CURRENT MICROBIOLOGY no. 67, edition 2, pages 240 - 248,
ISSN: 0343-8651
Language:
English
Publication year:
2013
Bibliographic description:
Kowal P., Ciesielski S.: Ribosomal intergenic spacer analysis as a tool for monitoring methanogenic archaea changes in an anaerobic digester// CURRENT MICROBIOLOGY. -Vol. 67, iss. 2 (2013), s.240-248
DOI:
Digital Object Identifier (open in new tab) 10.1007/s00284-013-0353-2
Verified by:
Gdańsk University of Technology

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