Journal
BIORESOURCE TECHNOLOGY
Volume 163, Issue -, Pages 40-47Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.biortech.2014.04.008
Keywords
Clostridia-specific primers; PCR-DGGE; Biodiversity; Dark fermentation pit mud; Complex microbial ecosystem
Funding
- National High Technology Research and Development Program of China [2013AA102108]
- National Nature Science Foundation of China [31271921]
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In this study, a Clostridia-specific primer set SJ-F and SJ-R, based on the available 165 rRNA genes sequences from database, was successfully designed and authenticated by theoretical and experimental evaluations. It targeted 19 clostridial families and unclassified_Clostridia with different coverage rates. The specificity and universality of novel primer set was tested again using the dark fermentation pit mud (FPM). It was demonstrated that a total of 13 closest relatives including 12 species were affiliated with 7 clostridial genera, respectively. Compared to the well-accepted bacterial universal primer pair P2/P3, five unexpected clostridial genera including Roseburia, Tissierella, Sporanaerobacter, Alkalibacter and Halothermothrix present in the FPM were also revealed. Therefore, this study could provide a good alternative to investigate the clostridial diversity and monitor their population dynamics rapidly and efficiently in various anaerobic environments and dark fermentation systems in future. (c) 2014 Elsevier Ltd. All rights reserved.
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