Interactions Among Sulfate-Reducing and Purple Sulfur Bacteria in the Chemocline of Meromictic Lake Cadagno, Switzerland
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Diss. ETH No. 15015 Interactions among sulfate-reducing and purple sulfur bacteria in the chemocline of meromictic Lake Cadagno, Switzerland A dissertation submitted to the SWISS FEDERAL INSTITUTE OF TECHNOLOGY ZURICH for the degree of DOCTOR OF NATURAL SCIENCES presented by SANDRO PEDUZZI Ing. dipl. EPFL born on October 15, 1973 from Airolo (Ticino) accepted on the recommendation of Prof. Dr. Alexander J.B. Zehnder, examiner Prof. Dr. Dittmar Hahn, co-examiner Dr. Mauro Tonolla, co-examiner 2003 Contents Summary....................................................................................................................................7 Résumé.......................................................................................................................................9 Chapter 1 13 Introduction 13 1.1 Stratified lakes........................................................................................................................... 14 1.2 Lake Cadagno............................................................................................................................ 17 1.3 The sulfur cycle......................................................................................................................... 20 1.3.1 Sulfate-reduction............................................................................................................... 24 1.3.2 Sulfur disproportionation .................................................................................................. 25 1.3.3 Anoxygenic photosynthesis .............................................................................................. 26 1.3.4 Sulfate-reducing and phototropic sulfur bacteria in Lake Cadagno.................................. 29 1.3 Aim of the thesis ....................................................................................................................... 32 1.4 References................................................................................................................................. 34 Chapter 2 45 In situ analysis of sulfate-reducing bacteria related to Desulfocapsa thiozymogenes in the chemocline of meromictic Lake Cadagno (Switzerland) 45 Abstract ........................................................................................................................................... 46 References....................................................................................................................................... 53 Chapter 3 57 Spatio-temporal distribution of phototrophic sulfur bacteria in the chemocline of meromictic Lake Cadagno (Switzerland) 57 3.1 Abstract ..................................................................................................................................... 58 3.2 Introduction............................................................................................................................... 58 3.3. Material and Methods............................................................................................................... 59 3.3.1. Site description, physical analyses and sampling............................................................. 59 3.3.2. Chemical analysis ............................................................................................................ 60 3.3.3. Microbial analysis............................................................................................................ 60 3.4 Results....................................................................................................................................... 62 3.4.1. Analysis of physico-chemical conditions......................................................................... 62 3.4.2. Microbial analysis............................................................................................................ 64 3.5 Discussion ................................................................................................................................. 68 3.6 References................................................................................................................................. 71 Chapter 4 77 Vertical distribution of sulfate-reducing bacteria in the chemocline of Lake Cadagno, Switzerland, over an annual cycle 77 Abstract ........................................................................................................................................... 78 References....................................................................................................................................... 89 Chapter 5 93 Isolation and characterization of aggregate-forming sulfate-reducing and purple sulfur bacteria from the chemocline of meromictic Lake Cadagno, Switzerland 93 5.1 Abstract ..................................................................................................................................... 94 5.2 Introduction............................................................................................................................... 94 5.3 Material and methods................................................................................................................ 96 5.3.1 Enrichment and isolation ..................................................................................................96 5.3.2 Identification and characterization.................................................................................... 97 5.3.3 Mixed culture study .......................................................................................................... 98 5.4 Results..................................................................................................................................... 100 5.4.1 Isolation and characterization of the sulfate-reducing bacterium, isolate Cad626.......... 100 5.4.2 Isolation and characterization of the phototrophic sulfur bacterium, isolate Cad16....... 101 5.4.3 Mixed culture study ........................................................................................................ 105 5.5 Discussion ............................................................................................................................... 106 5.6 References............................................................................................................................... 109 Chapter 6 115 Discussion 115 6.1 Aggregate formation ............................................................................................................... 116 6.2 Interactions among organisms involved in the sulfur cycle .................................................... 120 6.3 Interactions between sulfate-reducing and small-celled purple sulfur bacteria ...................... 124 6.4 Future perspectives.................................................................................................................. 131 6.5 References............................................................................................................................... 133 Curriculum vitae......................................................................................................................141 Aknowledgements...................................................................................................................143 Summary 7 Summary Permanently stratified lakes represent optimal model systems for the study of aquatic microorganisms since defined and stable vertical gradients of environmental conditions such as light intensity and quality, oxygen availability, or the presence of sulfide support the development of diverse populations of microorganisms adapted to defined ecological niches. Lake Cadagno is a meromictic lake located 1923 m above sea level in the Southern Alps of Switzerland (46°33'N, 8°43'E), in the catchment area of a dolomite vein rich in gypsum (Piora-Mulde). A permanent chemocline at a depth between 9 and 14 meters is stabilized by density differences of salt-rich water constantly supplied by subaquatic springs to the monimolimnion and of electrolyte-poor surface water feeding the mixolimnion. High concentrations of sulfate and steep sulfide gradients in the chemocline support the growth of large numbers of bacteria (up to 107 cells ml-1) indicating that a bacterial community making use of these gradients is present. Molecular techniques that were used to analyze microbial community structure unaffected by the limitations of culturability showed that almost all bacteria in the chemocline of Lake Cadagno belonged to the Proteobacteria with numbers for the -, -, - and -subdivision of Proteobacteria accounting for 23, 17, 45 and 15% of the total number of bacteria, respectively. Most prominent numerically (ca. 35% of all bacteria) were large- and small-celled phototrophic purple sulfur bacteria. The large-celled phototrophic sulfur bacteria were identified as Chromatium okenii, while small- celled phototrophic sulfur bacteria consisted of four major populations forming a tight cluster with Lamprocystis purpurea (former Amoebobacter purpureus and Pfennigia purpurea) and L. roseopersicina. The numerically most important phototrophic sulfur bacteria in the chemocline were small-celled purple sulfur bacteria of two yet uncultured populations designated D and F. Other small-celled purple sulfur bacteria (L. purpurea and L. roseopersicina) were found in numbers about one order of magnitude lower. These numbers were similar to those of the large-celled purple sulfur bacteria (C. okenii) and green