Abundance, Diversity and Activity of Sulfate-Reducing Prokaryotes in Heavy Metal-Contaminated Sediment from a Salt Marsh in the Medway Estuary (UK)

被引:35
作者
Quillet, Laurent [1 ]
Besaury, Ludovic [1 ]
Popova, Milka [1 ]
Paisse, Sandrine [1 ]
Deloffre, Julien [1 ]
Ouddane, Baghdad [2 ]
机构
[1] Univ Rouen, Fac Sci, Microbiol Grp, CNRS 6143 M2C, F-76821 Mont St Aignan, France
[2] Univ Sci & Technol Lille 1, UMR CNRS 8157, Equipe Chim Analyt & Marine, Lab Geosyst, F-59655 Villeneuve Dascq, France
关键词
Estuarine Sediment; Active Sulfate-reducing Prokaryotes; DNA Extraction Efficiency; dsrA Transcripts; SSCP; Phylogenetic Analysis; Heavy Metal Contamination; 16S RIBOSOMAL-RNA; SULFITE REDUCTASE GENES; GRADIENT GEL-ELECTROPHORESIS; IN-SITU HYBRIDIZATION; DESULFOVIBRIO-VULGARIS HILDENBOROUGH; 2 CONTRASTING MUDFLATS; FRESH-WATER SEDIMENT; WADDEN SEA SEDIMENTS; ACID-MINE DRAINAGE; REAL-TIME PCR;
D O I
10.1007/s10126-011-9420-5
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We investigated the diversity and activity of sulfate-reducing prokaryotes (SRP) in a 3.5-m sediment core taken from a heavy metal-contaminated site in the Medway Estuary, UK. The abundance of SRPs was quantified by qPCR of the dissimilatory sulfite reductase gene beta-subunit (dsrB) and taking into account DNA extraction efficiency. This showed that SRPs were abundant throughout the core with maximum values in the top 50 cm of the sediment core making up 22.4% of the total bacterial community and were 13.6% at 250 cm deep. Gene libraries for dsrA (dissimilatory sulfite reductase alpha-subunit) were constructed from the heavily contaminated (heavy metals) surface sediment (top 20 cm) and from the less contaminated and sulfate-depleted, deeper zone (250 cm). Certain cloned sequences were similar to dsrA found in members of the Syntrophobacteraceae, Desulfobacteraceae and Desulfovibrionaceae as well as a large fraction (60%) of novel sequences that formed a deep branching dsrA lineage. Phylogenetic analysis of metabolically active SRPs was performed by reverse transcription PCR and single strand conformational polymorphism analysis (RT-PCR-SSCP) of dsrA genes derived from extracted sediment RNA. Subsequent comparative sequence analysis of excised SSCP bands revealed a high transcriptional activity of dsrA belonging to Desulfovibrio species in the surface sediment. These results may suggest that members of the Desulfovibrionaceae are more active than other SRP groups in heavy metal-contaminated surface sediments.
引用
收藏
页码:363 / 381
页数:19
相关论文
共 123 条
[1]   Rapid detection and quantification of bisulfite reductase genes in oil field samples using real-time PCR [J].
Agrawal, Akhil ;
Lal, Banwari .
FEMS MICROBIOLOGY ECOLOGY, 2009, 69 (02) :301-312
[2]   Desulfovibrio hydrothermalis sp nov., a novel sulfate-reducing bacterium isolated from hydrothermal vents [J].
Alazard, D ;
Dukan, S ;
Urios, A ;
Verhé, F ;
Bouabida, N ;
Morel, F ;
Thomas, P ;
Garcia, JL ;
Ollivier, B .
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2003, 53 :173-178
[3]   BASIC LOCAL ALIGNMENT SEARCH TOOL [J].
ALTSCHUL, SF ;
GISH, W ;
MILLER, W ;
MYERS, EW ;
LIPMAN, DJ .
JOURNAL OF MOLECULAR BIOLOGY, 1990, 215 (03) :403-410
[4]   Single-cell identification in microbial communities by improved fluorescence in situ hybridization techniques [J].
Amann, Rudolf ;
Fuchs, Bernhard M. .
NATURE REVIEWS MICROBIOLOGY, 2008, 6 (05) :339-348
[5]   Effects of legacy nuclear waste on the compositional diversity and distributions of sulfate-reducing bacteria in a terrestrial subsurface aquifer [J].
Bagwell, CE ;
Liu, X ;
Wu, L ;
Zhou, J .
FEMS MICROBIOLOGY ECOLOGY, 2006, 55 (03) :424-431
[6]   FAST AND SENSITIVE SILVER STAINING OF DNA IN POLYACRYLAMIDE GELS [J].
BASSAM, BJ ;
CAETANOANOLLES, G ;
GRESSHOFF, PM .
ANALYTICAL BIOCHEMISTRY, 1991, 196 (01) :80-83
[7]   Chemical speciation of sulfur compounds in surface sediments from three bays (Fresnaye, Seine and Authie) in northern France, and identification of some factors controlling their generation [J].
Billon, G ;
Ouddane, B ;
Boughriet, A .
TALANTA, 2001, 53 (05) :971-981
[8]  
Boyle AW, 1999, APPL ENVIRON MICROB, V65, P1133
[9]  
Brown SD, 2011, J BACTERIOL IN PRESS, DOI 10.1128
[10]   A strategy for optimizing quality and quantity of DNA extracted from soil [J].
Bürgmann, H ;
Pesaro, M ;
Widmer, F ;
Zeyer, J .
JOURNAL OF MICROBIOLOGICAL METHODS, 2001, 45 (01) :7-20