This review of anaerobic metabolism emphasizes aerobic oxidation, because the two processes cannot be separated in a complete treatment of the topic. It is process oriented and highlights the fascinating microorganisms that mediate anaerobic biogeochemistry. We begin this review with a brief discussion of CO 2 assimilation by autotrophs, the source of most of the reducing power on Earth, and then consider the biological processes that harness this potential energy. Energy liberation begins with the decomposition of organic macromolecules to relatively simple compounds, which are simplified further by fermentation. Methanogenesis is considered next because CH 4 is a product of acetate fermentation, and thus completes the catabolism of organic matter, particularly in the absence of inorganic electron acceptors. Finally, the organisms that use nitrogen, manganese, iron, and sulfur for terminal electron acceptors are considered in order of decreasing free-energy yield of the reactions.

Anaerobic Metabolism: Linkages to Trace Gases and Aerobic Processes

The microbial assemblages of Lake Ciso and Lake Vilar (Banyoles, northeast Spain) were analyzed in space and time by microscopy and by performing PCR-denaturing gradient gel electrophoresis (DGGE) and sequence analysis of 16S rRNA gene fragments. Samples obtained from different water depths and at two different times of the year (in the winter during holomixis and in the early spring during a phytoplankton bloom) were analyzed. Although the lakes have the same climatic conditions and the same water source, the limnological parameters were different, as were most of the morphologically distinguishable photosynthetic bacteria enumerated by microscopy. The phylogenetic affiliations of the predominant DGGE bands were inferred by performing a comparative 16S rRNA sequence analysis. Sequences obtained from Lake Ciso samples were related to gram-positive bacteria and to members of the division Proteobacteria. Sequences obtained from Lake Vilar samples were related to members of the Cytophaga-Flavobacterium-Bacteroides phylum and to cyanobacteria. Thus, we found that like the previously reported differences between morphologically distinct inhabitants of the two lakes, there were also differences among the community members whose morphologies did not differ conspicuously. The changes in the species composition from winter to spring were also marked. The two lakes both contained sequences belonging to phototrophic green sulfur bacteria, which is consistent with microscopic observations, but these sequences were different from the sequences of cultured strains previously isolated from the lakes. Euryarchaeal sequences (i.e., methanogen- and thermoplasma-related sequences) also were present in both lakes. These euryarchaeal group sequences dominated the archaeal sequences in Lake Ciso but not in Lake Vilar. In Lake Vilar, a new planktonic population related to the crenarchaeota produced the dominant archaeal band. The phylogenetic analysis indicated that new bacterial and archaeal lineages were present and that the microbial diversity of these assemblages was greater than previously known. We evaluated the correspondence between the abundances of several morphotypes and DGGE bands by comparing microscopy and sequencing results. Our data provide evidence that the sequences obtained from the DGGE fingerprints correspond to the microorganisms that are actually present at higher concentrations in the natural system.

Identification of and Spatio-Temporal Differences between Microbial Assemblages from Two Neighboring Sulfurous Lakes: Comparison by Microscopy and Denaturing Gradient Gel Electrophoresis

Phototrophic sulfur bacteria often form mass developments in aquatic environments, either planktonic or benthic, where anoxic layers containing reduced sulfur compounds are exposed to light. This chapter summarizes a number of reports from the literature, collecting the information on the abundance of these bacteria as well as on their contribution to primary production. From the point of view of population dynamics, the abundance of these organisms is the consequence of a certain balance between growth and losses. Both specific growth rates, and specific rates of loss through several processes are analyzed in several environments, in an attempt to generalize on the growth status of blooms of phototrophic sulfur bacteria. The information available indicates the existence of an upper limit for the production of these bacteria in nature, and seems to suggest the existence of an upper limit for biomass based in the balance between growth and losses.

Ecology of Phototrophic Sulfur Bacteria

The biomass, phylogenetic composition, and photoautotrophic metabolism of green sulfur bacteria in the Black Sea was assessed in situ and in laboratory enrichments. In the center of the western basin, bacteriochlorophyll e (BChl e) was detected between depths of 90 and 120 m and reached maxima of 54 and 68 ng liter−1. High-pressure liquid chromatography analysis revealed a dominance of farnesyl esters and the presence of four unusual geranyl ester homologs of BChl e. Only traces of BChl e (8 ng liter−1) were found at the northwestern slope of the Black Sea basin, where the chemocline was positioned at a significantly greater depth of 140 m. Stable carbon isotope fractionation values of farnesol indicated an autotrophic growth mode of the green sulfur bacteria. For the first time, light intensities in the Black Sea chemocline were determined employing an integrating quantum meter, which yielded maximum values between 0.0022 and 0.00075 μmol quanta m−2 s−1 at the top of the green sulfur bacterial layer around solar noon in December. These values represent by far the lowest values reported for any habitat of photosynthetic organisms. Only one 16S rRNA gene sequence type was detected in the chemocline using PCR primers specific for green sulfur bacteria. This previously unknown phylotype groups with the marine cluster of the Chlorobiaceae and was successfully enriched in a mineral medium containing sulfide, dithionite, and freshly prepared yeast extract. Under precisely controlled laboratory conditions, the enriched green sulfur bacterium proved to be capable of exploiting light intensities as low as 0.015 μmol quanta m−2 s−1 for photosynthetic 14CO2 fixation. Calculated in situ doubling times of the green sulfur bacterium range between 3.1 and 26 years depending on the season, and anoxygenic photosynthesis contributes only 0.002 to 0.01% to total sulfide oxidation in the chemocline. The stable population of green sulfur bacteria in the Black Sea chemocline thus represents the most extremely low-light-adapted and slowest-growing type of phototroph known to date.

Physiology and Phylogeny of Green Sulfur Bacteria Forming a Monospecific Phototrophic Assemblage at a Depth of 100 Meters in the Black Sea

Contents: Introduction.- The Genus Azospirillum.- Other Root-Associated Diazotrophs.- Association with Plants.- Quantification of N2 Fixation.- Agronomic Implications.- Potentials and Prospects for the Future.- References.- Index.

Nitrogen-fixing bacteria in nonleguminous crop plants

The anaerobic zones of Lakes Cis6 and Vilar (Banyoles karstic area, NE Spain) had mass developments of purple sulfur bacteria during summer 1982. In Lake Vilar, Chromatium spp. was dominant (up to 92% of the microbial biovolume). In Lake Ciso, the predominant microorganisms were Chromatium spp. (up to 7 1%) and another purple sulfur bacterium forming aggregates (20%). The bacterial layer could be divided according to the physiological state of the cells into a top part of maximal specific activity, a peak of maximal abundance, and a bottom part of inactive cells. The bacteria in the peak were predominantly limited by light; sulfide, phosphate, and acetate were not limiting in the middle of the day. The light limitation started at the depth having the maximal concentration of cells; the top of the layer appeared to be sulfide-limited. Specific contents of photopigments, elemental sulfur, and reserve polymers decreased from the top to the bottom of the bacterial layer. These phenomena point to the crucial role of light in the development of layers of phototrophic bacteria in stratified lakes.

https://aslopubs.onlinelibrary.wiley.com/doi/pdf/10.4319/lo.1985.30.5.0919

Phototrophic sulfur bacteria in two Spanish lakes: Vertical distribution and limiting factors1

The effect of different concentrations of sulfide and sulfur on the assimilation of acetate by Chlorobium phaeobacteroides was investigated in batch and continuous cultures. In batch cultures the assimilation of acetate strictly depends on the initial concentration of sulfide. In continuous cultures the uptake of acetate depends not only on the reservoir concentration of sulfide but also on the dilution rate. The more severe the limitation of sulfide the higher the incorporation of acetate. The very efficient uptake of acetate was also observed in batch cultures, but only immediately prior to sulfide depletion. After sulfide depletion, with sulfur still available, the uptake of acetate per mmol reducing power increased even further. This phenomenon, which has been overlooked since growth decreases drastically after sulfide depletion due to incapacity for assimilatory sulfate reduction is of ecological importance in the formation of blooms of brown Chlorobium species.

P. A. G. Hofman

Papers

Phototrophic sulfur bacteria in two Spanish lakes: Vertical distribution and limiting factors1

Ecological significance of acetate assimilation by Chlorobium phaeobacteroides