Characterization and in situ carbon metabolism of phototrophic consortia

A dense population of the phototrophic consortium "Pelochromatium roseum" was investigated in the chemocline of a temperate holomictic lake (Lake Dagow, Brandenburg, Germany). Fluorescence in situ hybridization revealed that the brown epibionts of "P. roseum" constituted up to 37% of the total bacterial cell number and up to 88% of all green sulfur bacteria present in the chemocline. Specific amplification of 16S rRNA gene fragments of green sulfur bacteria and denaturing gradient gel electrophoresis fingerprinting yielded a maximum of four different DNA bands depending on the year of study, indicating that the diversity of green sulfur bacteria was low. The 465-bp 16S rRNA gene sequence of the epibiont of "P. roseum" was obtained after sorting of individual consortia by micromanipulation, followed by a highly sensitive PCR. The sequence obtained represents a new phylotype within the radiation of green sulfur bacteria. Maximum light-dependent H(14)CO(3)(-) fixation in the chemocline in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea suggested that there was anaerobic autotrophic growth of the green sulfur bacteria. The metabolism of the epibionts was further studied by determining stable carbon isotope ratios (delta(13)C) of their specific biomarkers. Analysis of photosynthetic pigments by high-performance liquid chromatography revealed the presence of high concentrations of bacteriochlorophyll (BChl) e and smaller amounts of BChl a and d and chlorophyll a in the chemocline. Unexpectedly, isorenieratene and beta-isorenieratene, carotenoids typical of other brown members of the green sulfur bacteria, were absent. Instead, four different esterifying alcohols of BChl e were isolated as biomarkers of green sulfur bacterial epibionts, and their delta(13)C values were determined. Farnesol, tetradecanol, hexadecanol, and hexadecenol all were significantly enriched in (13)C compared to bulk dissolved and particulate organic carbon and compared to the biomarkers of purple sulfur bacteria. The difference between the delta(13)C values of farnesol, the major esterifying alcohol of BChl e, and CO(2) was -7.1%, which provides clear evidence that the mode of growth of the green sulfur bacterial epibionts of "P. roseum" in situ is photoautotrophic.     

Quinones of phototrophic purple bacteria

Abstract The quinone composition of the recognized species of the phototrophic purple nonsulfur bacteria, the Ectothiorhodospiraceae, and some Chromatiaceae species has been determined. Altogether more than 50 strains of 33 species have been investigated. Some of the purple nonsulfur bacteria have Q-10 as sole quinone component, while others have Q-10, Q-9, or Q-8, respectively, together with menaquinones of the same isoprenoid chain length as the major components. Rhodoquinone is present in Rhodospirillum rubrum and Rhodospirillum photometricum . The Ectothiorhodospira species have either Q-8 and MK-8, like the Chromatiaceae species, or Q-7 and MK-7 as the major components.     

The influence that protozoa has on hydrogen sulfide oxidation by phototrophic bacteria

These experiments have been performed with a facility for the simulation of summer stagnation conditions in natural water bodies. Our experimental results suggest that anaerobic ciliates and flagellates can stimulate hydrogen sulfide oxidation and thus the photosynthesis of phototrophic green bacteria in phosphorus deficient media. The phosphorus (as well as nitrogen) limitation of hydrobiological processes is most widespread in natural waters and, therefore, our results may contribute to the understanding of the role that individual organisms and their relationships play in aquatic ecosystems.     

Biogeography, evolution, and diversity of epibionts in phototrophic consortia

Motile phototrophic consortia are highly regular associations in which numerous cells of green sulfur bacteria surround a flagellated colorless beta-proteobacterium in the center. To date, seven different morphological types of such consortia have been described. In addition, two immotile associations involving green sulfur bacteria are known. By employing a culture-independent approach, different types of phototrophic consortia were mechanically isolated by micromanipulation from 14 freshwater environments, and partial 16S rRNA gene sequences of the green sulfur bacterial epibionts were determined. In the majority of the lakes investigated, different types of phototrophic consortia were found to co-occur. In all cases, phototrophic consortia with the same morphology from the same habitat contained only a single epibiont phylotype. However, morphologically indistinguishable phototrophic consortia collected from different lakes contained different epibionts. Overall, 19 different types of epibionts were detected in the present study. Whereas the epibionts within one geographic region were very similar (Dice coefficient, 0.582), only two types of epibionts were found to occur on both the European and North American continents (Dice coefficient, 0.190). None of the epibiont 16S rRNA gene sequences have been detected so far in free-living green sulfur bacteria, suggesting that the interaction between epibionts and chemotrophic bacteria in the phototrophic consortia is an obligate interaction. Based on our phylogenetic analysis, the epibiont sequences are not monophyletic. Thus, the ability to form symbiotic associations either arose independently from different ancestors or was present in a common ancestor prior to the radiation of green sulfur bacteria and the transition to the free-living state in independent lineages. The present study thus demonstrates that there is great diversity and nonrandom geographical distribution of phototrophic consortia in the natural environment.     

Active transport in phototrophic bacteria

Phototrophic bacteria utilize light-driven, cyclic electron flow to pump protons out of their cytoplasm, creating an electrochemical proton gradient, _H+, outside acid and positive. These bacteria exchange external protons for internal cations (Na⁺, K⁺ and Ca⁺²), allowing the cells to maintain a nearly constant internal pH while maintaining the electrical component of _H+. Na⁺/H⁺ exchange also establishes an electrochemical Na⁺ gradient. Phototrophic bacteria are able to utilize these electrochemical gradients as energy sources for the uptake of a wide variety of metabolites (e.g., sugars, organic acids and amino acids) via metabolite/cation symports.     

Organic nitrogen metabolism of phototrophic bacteria

Recent reviews dealing with phototrophic bacteria are concerned with bioenergetics, nitrogen fixation and hydrogen metabolism, synthesis of the photosynthetic apparatus and phylogeny/taxonomy. The organic N-metabolism of these phylogenetically diverse bacteria has last been reviewed in 1978. However, amino acid utilization and biosynthesis, ammonia assimilation, purine and pyrimidine metabolism and biosynthesis of -aminolevulinic acid as precursor of bacteriochlorophylls and hemes are topics of vital importance. This review focusses on utilization of amino acids as N- and C/N-sources, the pathways of purine and pyrimidine degradation, novel aspects of amino acid biosynthesis (with emphasis on branched-chain amino acids and -aminole-vulinic acid) and some aspects of ammonia assimilation and glutamate synthesis by purple bacteria, green sulfur bacteria and Chloroflexus aurantiacus.Abbreviations R Rhodospirillum - Rhb Rhodobacter - Rc Rhodocyclus - Rp Rhodopila - Rps Rhodopseudomonas     

Phylogenetic affiliations ofRhodoferax fermentans and related species of phototrophic bacteria as determined by automated 16S rDNA sequencing

16S rDNA sequences of strains ofRhodoferax fermentans were analyzed and compared with those of species of the generaRubrivivax andRhodocyclus. Approximately 1.5-kb fragments of 16S rDNA from crude cell lysates were amplified by the polymerase chain reaction (PCR) and sequenced directly by usingTth DNA polymerase with the linear PCR sequencing protocol, followed by on-line detection with an automated laser fluorescent DNA sequencer. Pairwise sequence comparisons and distance matrix tree analysis showed thatRhodoferax fermentans, Rubrivivax gelatinosus, andRhodocyclus species belong to three separate lineages within the beta subclass of theProteobacteria, thereby confirming the phylogenetic validity of the genusRhodoferax, as well as of the generaRubrivivax andRhodocyclus.     

Anoxygenic phototrophic bacteria from extreme environments

A diverse group of anoxygenic phototrophic bacteria thrive in habitats characterized by extremes of temperature, pH, or salinity. These `extremophilic' anoxygenic phototrophs are optimally adapted to the conditions of their habitats and are ideal model systems for defining the physiochemical limits of photosynthesis. Extremophilic phototrophs have provided new insight into the evolution of photosynthesis and play ecological roles as primary producers in their unusual habitats. This revised version was published online in August 2006 with corrections to the Cover Date.     

Viable phototrophic sulfur bacteria from the Black-Sea bottom

Phototrophic sulfur bacteria were isolated from Black-Sea mud at depths of 660 and 2,240 m. The species obtained in pure cultures were identified asChromatium warmingii andThiocapsa roseopersicina. In addition,Chlorobium phaeovibrioides occurred in enrichment cultures. The findings prove that Chromatiaceae and Chlorobiaceae are capable of survival in 2000 m depth, in the dark, in the presence of H₂S and organic materials. There is, however, no evidence for growth or reproduction of these organisms in this environment.     

A modified method for the cultivation of phototrophic bacteria

Simplified anaerobic media and a convenient method for the cultivation of Rhodospirillaceae, Chlorobiaceae, Chloroflexaceae and Chromatiaceae are described. The modified conditions assure almost complete anaerobiosis for media, growth and maintenance.Strains representing several species of Rhodospirillaceae, Chlorobiaceae and Chromatiaceae were successfully grown within relatively short times with full pigmentation, indicating that the new media and cultivation conditions were most suited for photoautotrophic growth.     

Interactions between phototrophic bacteria in sediment ecosystems

The environmental conditions in laminated microbial sediment ecosystems on the island of Schiermonnikoog (The Netherlands) were monitoredin situ over 24-hour periods by using micro-electrodes. In the layer of purple sulfur bacteria dramatic diel aerobic/anaerobic shifts occurred, whereas the top layer of cyanobacteria was occasionally confronted with sulfide. Pure cultures of the dominant organisms, being the cyanobacteriumMicrocoleus chthonoplastes and the purple sulfur bacteriumThiocapsa roseopersicina, were subjected to regimes mimicking the natural circumstances. It was demonstrated that both organisms are physiologically very well adapted to the fluctuating environmental conditions. The organisms interact by releasing metabolic end-products, the removal of toxic compounds and by competition for common substrates. It was demonstrated that positive interactions between both organisms are more important than negative interactions.     

Cysteine and S-sulfocysteine biosynthesis in phototrophic bacteria

Forteen species (17 strains) of phototrophic bacteria as well as one strain of Thiobacillus denitrificans were tested for cysteine synthase and S-sulfocysteine synthase. All strains contain cysteine synthase active with O-acetylserine; only the Chromatiaceae, two species of the Rhodospirillaceae and T. denitrificans contain S-sulfocysteine synthase. In six species repression by different sulfur compounds in the medium was studied. In Chromatium vinosum, cysteine synthase was found to be constitutive, while in the Rhodospirillaceae tested the enzyme is repressed by sulfide. Thiosulfate had a derepressive effect in Rhodopseudomonas globiformis but strongly repressed cysteine synthase in R. sulfidophila and R. palustris. Cysteine had only moderate effects with the species tested.     

True marine and halophilic anoxygenic phototrophic bacteria

Anoxygenic phototrophic bacteria are widely distributed in marine sediments and shallow waters of the coastal zone, where they often form intensely colored mass developments. The phototrophic bacteria have adapted to the whole spectrum of salt concentrations, from freshwater to saturated brines, and it is apparent that individual species have adapted well to particular habitats and mineral salts compositions, both qualitatively and quantitatively. This adaptation is reflected not only in the demand for defined ranges of salt concentrations, but also in the phylogenetic relationships of these bacteria, as established by 16S rDNA sequences. Major phylogenetic branches of purple sulfur bacteria are represented by: (1) marine and extremely halophilic Ectothiorhodospiraceae, (2) truly marine and halophilic Chromatiaceae and (3) freshwater Chromatiaceae, some of which are tolerant to low salt concentrations and are successful competitors in brackish and marine habitats. Quite similarly, salt-dependent green sulfur bacteria form distinct phylogenetic lines. In addition, also among the phototrophic alpha-Proteobacteria (purple nonsulfur bacteria), distinct phylogenetic lines of salt-dependent species are recognized. Available data give rise to the assumption that salt concentrations of natural habitats are an important selective factor that determines the development of a selected range of phototrophic bacteria in an exclusive way. As a consequence, the salt responses of these bacteria are reflected in their phylogenetic relationships.     

Marine sponges as habitats of anaerobic phototrophic bacteria

Enrichment cultures were prepared with different media for phototrophic bacteria from four species of marine sponges, collected from oxic coastal waters near Split (Yugoslavia). We obtained pure cultures of six strains ofChromatiaceae and two strains ofRhodospirillaceae by agar shake dilution. TheRhodospirillaceae were identified asRhodopseudomonas sulfidophila and a marine form ofRhodopseudomonas palustris. TheChromatiaceae were identified asChromatium vinosum, Chromatium gracile, Chromatium minutissimum. Ectothiorhodospira mobilis, and a Chromatium species, which in some respects resemblesChromatium minus. The occurrence of strictly anaerobic phototrophic bacteria in aerobic sponges is discussed with respect to nutrition and possible syntrophism.