Purple Sulfur Research Articles

Utilization of solid ‘elemental’ sulfur by the phototrophic purple sulfur bacterium Allochromatium vinosum: a sulfur K-edge X-ray absorption spectroscopy study

The purple sulfur bacterium Allochromatium vinosum can use elemental sulfur as an electron donor for anoxygenic photosynthesis. The elemental sulfur is taken up, transformed into intracellular sulfur globules and oxidized to sulfate. Commercially available "elemental" sulfur usually consists of the two species cyclo-octasulfur and polymeric sulfur. The authors investigated whether only one sulfur species is used or at least preferred when Alc. vinosum takes up elemental sulfur and forms globules. To this end, Alc. vinosum was cultivated photolithoautotrophically with two types of elemental sulfur that differed in their cyclo-octasulfur : polymeric sulfur ratio, as well as with pure polymeric sulfur. Sulfur speciation was analysed using X-ray absorption spectroscopy, and sulfate contents were determined by HPLC to quantify the amount of elemental sulfur being taken up and oxidized by Alc. vinosum. The results show that Alc. vinosum uses only the polymeric sulfur (sulfur chain) fraction of elemental sulfur and is probably unable to take up and form sulfur globules from cyclo-octasulfur. Furthermore, direct cell-sulfur contact appears to be necessary for uptake of elemental sulfur by Alc. vinosum.

The role of Hox hydrogenase in the H 2 metabolism of Thiocapsa roseopersicina

The purple sulfur phototrophic bacterium Thiocapsa roseopersicina BBS synthesizes at least three NiFe hydrogenases (Hox, Hup, Hyn). We characterized the physiological H 2 consumption/evolution reactions in mutants having deletions of the structural genes of two hydrogenases in various combinations. This made possible the separation of the functionally distinct roles of the three hydrogenases. Data showed that Hox hydrogenase (unlike the Hup and Hyn hydrogenases) catalyzed the dark fermentative H 2 evolution and the light-dependent H 2 production in the presence of thiosulfate. Both Hox + and Hup + mutants demonstrated light-dependent H 2 uptake stimulated by CO 2 but only the Hup + mutant was able to mediate O 2-dependent H 2 consumption in the dark. The ability of the Hox + mutant to evolve or consume hydrogen was found to depend on a number of interplaying factors including both growth and reaction conditions (availability of glucose, sulfur compounds, CO 2, H 2, light). The study of the redox properties of Hox hydrogenase supported the reversibility of its action. Based on the results a scheme is suggested to describe the role of Hox hydrogenase in light-dependent and dark hydrogen metabolism in T. roseopersicina BBS.

Characterization of the endoevaporitic microbial communities in a hypersaline gypsum crust by fatty acid analysis

We have used fatty acid analyses to study the community structure of a layered endoevaporitic microbial community within a gypsum crust that covers the bottom of a saltern evaporation pond in Eilat, Israel. This community, living at a salinity of 218–238 g l−1 total dissolved salts, consists of an upper brown layer dominated by unicellular cyanobacteria, a green layer with filamentous cyanobacteria, a red-purple layer with both Chromatium and Ectothiorhodospira/Halorhodospira type of purple sulfur bacteria, and a black layer in which dissimilatory sulfate reduction occurs. An olive-green layer is sometimes present below the red-purple layer. Analysis by gas chromatography/mass spectrometry of the fatty acid methyl esters prepared from the different fractions showed characteristic patterns in each layer, and these could be related to fatty acid composition data from the literature and to fatty acid analyses of representative halophilic microorganisms isolated from the site. The nature of the fatty acids in the green layer suggests that the cyanobacteria present there use the oxygen-independent pathway for production of unsaturated fatty acids, a pathway only occasionally encountered in filamentous cyanobacteria. The facultative anaerobic nature of the cyanobacteria in the green layer was confirmed by their ability to perform anoxygenic photosynthesis with sulfide as electron donor. Specific signature fatty acids identified for each layer corresponded well with the microscopic and functional analysis of the biota present.

Thiosulphate oxidation in the phototrophic sulphur bacterium Allochromatium vinosum

Two different pathways for thiosulphate oxidation are present in the purple sulphur bacterium Allochromatium vinosum: oxidation to tetrathionate and complete oxidation to sulphate with obligatory formation of sulphur globules as intermediates. The tetrathionate:sulphate ratio is strongly pH-dependent with tetrathionate formation being preferred under acidic conditions. Thiosulphate dehydrogenase, a constitutively expressed monomeric 30 kDa c-type cytochrome with a pH optimum at pH 4.2 catalyses tetrathionate formation. A periplasmic thiosulphate-oxidizing multienzyme complex (Sox) has been described to be responsible for formation of sulphate from thiosulphate in chemotrophic and phototrophic sulphur oxidizers that do not form sulphur deposits. In the sulphur-storing A. vinosum we identified five sox genes in two independent loci (soxBXA and soxYZ). For SoxA a thiosulphate-dependent induction of expression, above a low constitutive level, was observed. Three sox-encoded proteins were purified: the heterodimeric c-type cytochrome SoxXA, the monomeric SoxB and the heterodimeric SoxYZ. Gene inactivation and complementation experiments proved these proteins to be indispensable for thiosulphate oxidation to sulphate. The intermediary formation of sulphur globules in A. vinosum appears to be related to the lack of soxCD genes, the products of which are proposed to oxidize SoxY-bound sulphane sulphur. In their absence the latter is instead transferred to growing sulphur globules.

Selective enrichment of green sulfur bacteria in the presence of 4-aminobenzenesulfonate (sulfanilate)

A novel selective enrichment method is described for phototrophic green sulfur bacteria even in the presence of purple sulfur and purple nonsulfur bacteria using sulfanilate, which was discovered during efforts to selectively isolate sulfanilate-metabolizing anoxygenic phototrophic bacteria from marine habitats. Samples for these experiments were obtained from beaches, saltpans, subsurface mangrove soils, fish and prawn aquaculture ponds and backwaters of the East and West coasts of India. Photoorganoheterotrophic and photolithoautotrophic enrichments in the absence of sulfanilate predominantly yielded purple bacterial enrichments. In contrast, photolithoautotrophic enrichments in the presence of sulfanilate yielded green-colored enrichments from the same samples. Whole cell absorption spectra of the enrichment cultures revealed the presence of bacteriochlorophyll c and thus green phototrophic bacteria. Microscopic observation demonstrated the presence of sulfur globules outside the bacterial cells and the presence of non-motile cells, some of which had prosthecae. 16S rDNA sequences obtained from green sulfur bacterial strains isolated from enrichment cultures confirmed the presence of representatives of the green sulfur bacterial genera Prosthecochloris and Chlorobaculum. The selective pressure of sulfanilate exerted through inhibition of phototrophic purple sulfur bacteria was demonstrated by inhibition studies using the purple sulfur bacteria Marichromatium indicum JA100 and Marichromatium sp. JA120 (JCM 13533) and the green sulfur bacterium Prosthecochloris sp. JAGS6 (JCM 13299).

Molecular and isotopic characterization of biomarkers in the Frick Swiss Jura sediments: A palaeoenvironmental reconstruction on the northern Tethys margin

Molecular and stable carbon isotope compositions of source-specific hydrocarbons have been used to reconstruct palaeoenvironmental conditions during deposition of the Middle Hettangian to Upper Sinemurian sediments on the northern epicontinental Tethys margin, Frick Swiss Jura. Increasing algal, cyanobacterial and phytoplanktonic (i.e., dinoflagellate) contributions associated with the 13C-enrichment of cyanobacteria derivatives (i.e., hopanes and monomethylalkanes) suggest enhanced primary productivity upsection. This is related to the 13C-enrichment of dissolved CO 2 in the upper layers and the progressive increase of depth and oxygenation of the water column. In the Middle Hettangian shallow-water environments (lagoon), the occurrence of green sulfur bacteria ( Chlorobiaceae) derivatives indicates that the lower part of the water column was strictly anoxic and rich in H 2S. Since these bacteria require very low light intensity to grow, these euxinic conditions may be extended up to the photic zone, allowing for anaerobic photosynthesis. Light penetration depth is most likely reduced by high productivity and/or turbidity in the photic zone. In these sediments, 13C-depleted hopanoids (−39.5‰) are most likely associated with phototrophic purple sulfur bacteria utilizing isotopically light organic carbon at the base of the aerobic zone. These purple sulfur bacteria may have consumed the H 2S used by Chlorobiaceae in the deeper layers and thus, sustained the algae and cyanobacteria productivity in the upper layers. The 13C-depleted carbonate (−13.3‰) may be partially related to the anaerobic oxidation of the organic matter during bacterial sulfate-reduction.

Crystallographic and X-ray diffraction analysis of metalloenzymes from purple sulphur bacterium

Crystallographic and X-ray diffraction analysis of metalloenzymes from purple sulphur bacterium

Diversity of phototrophic bacteria in microbial mats from Arctic hot springs (Greenland)

We investigated the genotypic diversity of oxygenic and anoxygenic phototrophic microorganisms in microbial mat samples collected from three hot spring localities on the east coast of Greenland. These hot springs harbour unique Arctic microbial ecosystems that have never been studied in detail before. Specific oligonucleotide primers for cyanobacteria, purple sulfur bacteria, green sulfur bacteria and Choroflexus/Roseiflexus-like green non-sulfur bacteria were used for the selective amplification of 16S rRNA gene fragments. Amplification products were separated by denaturing gradient gel electrophoresis (DGGE) and sequenced. In addition, several cyanobacteria were isolated from the mat samples, and classified morphologically and by 16S rRNA-based methods. The cyanobacterial 16S rRNA sequences obtained from DGGE represented a diverse, polyphyletic collection of cyanobacteria. The microbial mat communities were dominated by heterocystous and non-heterocystous filamentous cyanobacteria. Our results indicate that the cyanobacterial community composition in the samples were different for each sampling site. Different layers of the same heterogeneous mat often contained distinct and different communities of cyanobacteria. We observed a relationship between the cyanobacterial community composition and the in situ temperatures of different mat parts. The Greenland mats exhibited a low diversity of anoxygenic phototrophs as compared with other hot spring mats which is possibly related to the photochemical conditions within the mats resulting from the Arctic light regime.

Thiorhodococcus mannitoliphagus sp. nov., a purple sulfur bacterium from the White Sea

A novel purple sulfur bacterium, strain WS(T), was isolated from a microbial mat from an estuary of the White Sea. Individual cells are coccoid shaped, motile by flagella and do not contain gas vesicles. The mean cell diameter is 1.85 mum (range 1.5-2.0 mum). Cell suspensions exhibit a purple-violet colour. They contain bacteriochlorophyll a and carotenoids of the rhodopinal series as photosynthetic pigments. The novel bacterium is an anoxygenic photoautotroph, using sulfide, thiosulfate, sulfite and elemental sulfur as electron donors for photosynthesis and is capable of photoassimilating several organic carbon sources in the presence of carbonate and a reduced sulfur source (sulfide and/or thiosulfate). Sulfur globules, formed during oxidation of sulfide, are stored transiently inside the cells. Optimal salinity and pH for growth are at 0.5-2.0 % NaCl and pH 7.0-7.5. The DNA base composition of strain WS(T) is 61.8 mol% G+C. 16S rRNA gene sequence analysis showed that the new isolate belongs to the genus Thiorhodococcus, with Thiorhodococcus minor CE2203(T) as the nearest relative (sequence similarity of 97.3 %). Several distinct differences from described species necessitate the description of a novel species. Thiorhodococcus mannitoliphagus sp. nov. is the proposed name, with strain WS(T) (=ATCC BAA-1228(T)=VKM B-2393(T)) as the type strain.

Cross-crystallization method used for the crystallization and preliminary diffraction analysis of a novel di-haem cytochrome c4.

The newly discovered di-haem cytochrome c4 from the purple sulfur photosynthetic bacterium Thiocapsa roseopersicina is the first cytochrome c4 to be crystallized from an anaerobic organism. It was crystallized using the addition of metal-ion salts to the standard vapour-diffusion method. Coloured well shaped three-dimensional crystals with dimensions of approximately 0.6 x 0.05 x 0.02 mm grew within 3-4 d at pH 5 and diffracted to 1.72 angstroms without radiation damage. Cytochrome c4 crystallized in space group P4(1)2(1)2 as a primitive tetragonal system with unit-cell parameters a = b = 75.29, c = 37.12 angstroms, alpha = beta = gamma = 90 degrees.

A novel approach for biohydrogen production

H 2 is considered as the ultimate cleanest energy carrier to be generated from renewable sources. Biohydrogen production is carried out using the enzymes nitrogenase or hydrogenase. The purple sulphur phototrophic bacterium, Thiocapsa roseopersicina BBS contains a nitrogenase and several NiFe hydrogenases. Hydrogenases can be used as fuel cell H 2 splitting catalyst and nitrogenase(s) and hydrogenases in biological H 2 production. The membrane-associated Hyn and Hup hydrogenases preferentially work in the H 2 uptake direction in vivo, the cytoplasmic Hox enzyme is a bidirectional enzyme. Non-nitrogen-fixing conditions, which render Hup and Hyn non-functioning, lead to sustained net hydrogen production via the Hox enzyme. The cumulative H 2 production by the apparently constitutive Hox and the inducible nitrogenase may become comparable under appropriate conditions.

Ecology of Algal Mats from Hypersaline Ponds in the British Virgin Islands

Benthic sediment samples ranging from poorly aggregated sand to complex, stratified mats were collected from six hypersaline ponds from March and July 1995 in the British Virgin Islands. Assemblages were analyzed with respect to species composition and abundance within visibly distinct layers in each mat sample. In individual ponds there was no apparent association between changing depth and the development of the benthic mats. Some species were present in all samples (e.g. Oscillatoria sp.) while others were restricted to single sites (e.g. Johannesbaptistia pellucida). Primary species included Microcoleus chthonoplastes, Phormidium spp., Coccochloris stagnina, and purple sulfur bacteria. Quantitative analysis of community structure included cluster and principal component analysis. Samples from individual ponds were often clustered; however, this was subject to seasonal variation. Mats collected in March were generally thicker and contained more layers than those in July. Variation among sites was not explained by the measured variation in environmental factors such as average pond salinity, depth, and oxygen concentration (mg/L). This study provides a detailed analysis of mat communities in hypersaline ponds and compares them with similar mat communities from other areas.

Siro(haem)amide inAllochromatium vinosumand relevance of DsrL and DsrN, a homolog of cobyrinic acida,c-diamide synthase, for sulphur oxidation

In the purple sulphur bacterium Allochromatium vinosum, the prosthetic group of dissimilatory sulphite reductase (DsrAB) was identified as siroamide, an amidated form of the classical sirohaem. The genes dsrAB are the first two of a large cluster of genes necessary for the oxidation of sulphur globules stored intracellularly during growth on sulphide and thiosulphate. DsrN is homologous to cobyrinic acid a,c diamide synthase and may therefore catalyze glutamine-dependent amidation of sirohaem. Indeed, an A. vinosumDeltadsrN in frame deletion mutant showed a significantly reduced sulphur oxidation rate that was fully restored upon complementation with dsrN in trans. Sulphite reductase was still present in the DeltadsrN mutant. DsrL is a homolog of the small subunits of bacterial glutamate synthases and was proposed to deliver glutamine for sirohaem amidation. However, recombinant DsrL does not exhibit glutamate synthase activity nor does the gene complement a glutamate synthase-deficient Escherichia coli strain. Deletion of dsrL showed that the encoded protein is absolutely essential for sulphur oxidation in A. vinosum.

Biomarker analysis of microbial diversity in sediments of a saline groundwater seep of Salt Basin, Nebraska

Lipids extracted from sediments in a saline seep in the Salt Basin of Lancaster County, Nebraska included alkanes, alkenes, alkanols, phytol, C 27–30 sterols, C 30–32 hopanoids, tetrahymanol, glycolipid and phospholipid fatty acids, and lipopolysaccharide hydroxyl fatty acids. Biomarker profiles suggest that the brine seeps of Salt Basin support a microbial ecosystem adapted to a relatively highly saline and sulfidic environment. The phospholipid fatty acid (PLFA) and lipopolysaccharide hydroxyl fatty acid profiles are consistent with the presence of large numbers of sulfate-reducing bacteria (SRB) in black, sulfidic muds surrounding the seeps. In the context of field and laboratory observations, the presence of large amounts of glycolipid fatty acids is attributed to large populations of photosynthetic microorganisms (cyanobacteria, phytoplankton, and purple sulfur bacteria) that likely play important roles in the local cycling of carbon and sulfur. The sterol profile and the detection of polyunsaturated alkenes (C 21:6, C 21:7, C 30:4, and C 30:5) implicates microalgae as important contributors of organic matter at the site. Comparatively high concentrations of phytol (58.2 μg g −1 dry wt sediment) record the activity of photosynthetic organisms in the system. The δ 13C of phytol (−37.1‰) is compatible with a dominance of microalgae, cyanobacteria, or higher plants and a lesser contribution from phototrophic sulfur bacteria. The presence of various intermediate degradation products of phytol (phytenes and phytadienes) indicates that SRB likely mediate the chemical reduction of phytol in the anaerobic zone. The presence of C 30–32 hopanols can be attributed to cyanobacteria and methanotrophs in oxic regions of the water column, whereas bacterivorous ciliates and phototrophic sulfur bacteria living at the chemocline are likely sources of tetrahymanol. The carbon isotopic composition of individual fatty acids and neutral lipids helps to identify source organisms. These microorganisms and others constitute a unique and integrated ecosystem prescribed by the geochemistry of the Salt Basin.

GHP, a new c‐type green heme protein from Halochromatium salexigens and other proteobacteria

We have isolated a minor soluble green-colored heme protein (GHP) from the purple sulfur bacterium, Halochromatium salexigens, which contains a c-type heme. A similar protein has also been observed in the purple bacteria Allochromatium vinosum and Rhodopseudomonas cryptolactis. This protein has wavelength maxima at 355, 420, and 540 nm and remains unchanged upon addition of sodium dithionite or potassium ferricyanide, indicating either an unusually low or high redox potential, respectively. The amino-acid sequence indicates one heme per peptide chain of 72 residues and reveals weak similarity to the class I cytochromes. The usual sixth heme ligand methionine in these proteins appears to be replaced by a cysteine in GHP. Only one known cytochrome has a cysteine sixth ligand, SoxA (cytochrome c-551) from thiosulfate-oxidizing bacteria, which is low-spin and has a high redox potential because of an un-ionized ligand. The native size of GHP is 34 kDa, its subunit size is 11 kDa, and the net charge is -12, accounting for its very acidic nature. A database search of complete genome sequences reveals six homologs, all hypothetical proteins, from Oceanospirillum sp., Magnetococcus sp., Thiobacillus denitrificans, Dechloromonas aromatica, Thiomicrospira crunogena and Methylobium petroleophilum, with sequence identities of 35-64%. The genetic context is different for each species, although the gene for GHP is transcriptionally linked to several other genes in three out of the six species. These genes, coding for an RNAse, a protease/chaperone, a GTPase, and pterin-4a-carbinolamine dehydratase, appear to be functionally related to stress response and are linked in at least 10 species.

Comparison of cultivation‐dependent and molecular methods for studying the diversity of anoxygenic purple phototrophs in sediments of an eutrophic brackish lagoon

Phototrophic anoxygenic purple bacteria play a key role in many aquatic ecosystems by oxidizing sulfur compounds and low-molecular-weight organic compounds using light as energy source. In this study, molecular methods based upon pufM gene (photosynthetic unit forming gene) were compared with culture-dependent methods to investigate anoxygenic purple phototrophic communities in sediments of an eutrophic brackish lagoon. Thirteen strains, belonging to eight different genera of purple phototrophic bacteria were isolated with a large dominance of the metabolically versatile purple non-sulfur bacteria (eight strains), some purple sulfur bacteria (three strains) and two strains belonging to the Roseobacter clade (aerobic phototrophs). The pufM genes amplified from the isolated strains were not detected by the molecular methods [terminal-restriction fragment length polymorphism (T-RFLP)] applied on in situ communities. An environmental clone library of the pufM gene was thus constructed from sediment samples. The results showed that most of the clones probably corresponded to aerobic phototrophic bacteria. Our results demonstrate that the culture-dependent techniques remain the best experimental approach for determining the diversity of phototrophic purple non-sulfur bacteria whereas the molecular approach clearly illustrated the abundance of organisms related to the Roseobacter clade in these eutrophic sediments.

An outer membrane protein A (ompA) homologue from the photosynthetic purple sulfur bacterium Allochromatium vinosum

A 991 bp DNA fragment, consisting of a 225 amino acid reading frame homologous to outer membrane protein coding ompA gene, was cloned from a purple sulfur bacterium Allochromatium vinosum. The homology analysis revealed up to 51% similarity to other bacterial species. The absence of branching within diazotrophs or other taxonomically related groups shows the structural importance of the protein regardless of the metabolism and evolution of the species.

Cloning and characterization of nif structural and regulatory genes in the purple sulfur bacterium, Halorhodospira halophila

Halorhodospira halophila is a halophilic photosynthetic bacterium classified as a purple sulfur bacterium. We found that H. halophila generates hydrogen gas during photoautotrophic growth as a byproduct of a nitrogenase reaction. In order to consider the applied possibilities of this photobiological hydrogen generation, we cloned and characterized the structural and regulatory genes encoding the nitrogenase, nifH, nifD and nifA, from H. halophila. This is the first description of the nif genes for a purple sulfur bacterium. The amino-acid sequences of NifH and NifD indicated that these proteins are an Fe protein and a part of a MoFe protein, respectively. The important residues are conserved completely. The sequence upstream from the nifH region and sequence similarities of nifH and nifD with those of the other organisms suggest that the regulatory system might be a NifL-NifA system; however, H. halophila lacks nifL. The amino-acid sequence of H. halophila NifA is closer to that of the NifA of the NifL-NifA system than to that of NifA without NifL. H. halophila NifA does not conserve either the residue that interacts with NifL or the important residues involved in NifL-independent regulation. These results suggest the existence of yet another regulatory system, and that the development of functional systems and their molecular counterparts are not necessarily correlated throughout evolution. All of these Nif proteins of H. halophila possess an excess of acidic residues, which acts as a salt-resistant mechanism.

Morphological and ultrastructural characterization of an unusual purple sulfur bacterium from a marine microbial-mat community

An unusual purple sulfur bacterium present in the pink layer of the Ebro Delta microbial mats has been identified through the study of its ultrastructural features. As pure cultures of this bacterium have not been obtained, due to its inability to grow in axenic conditions, only enrichment cultures where it reached at least 90% of total biomass have been considered. In enrichment cultures, the cells are rods that are 5.4 ± 0.6 μm wide and 11.0 ± 2.1 μm long. The color of the cell suspensions is pink to pinkish-red. Cells are motile by means of a polar tuft of flagella and multiply by binary fission. This bacterium possesses an extensive internal photosynthetic membrane system consisting of stacks of lamellae, contains bacteriochlorophyll a and carotenoids of normal spirilloxanthin series and deposits sulfur intracellularly. In natural samples, the most abundant phototrophic purple sulfur bacterium developing in this ecosystem has the same kind of intracytoplasmic membrane system, but the cells differ slightly in size and arrangement, in that they are ovoid, 4.2 ± 0.5 μm wide and 6.0 ± 1.4 μm long, and can be seen forming irregular non-motile clumps which are embedded in slime. Differences observed between field samples and enrichment cultures suggest that environmental conditions may bring about changes in the phenotypic characteristics of the cells. The morphological characteristics of the described bacterium resemble those of large Chromatia. However, this bacterium differs from known species in this group, particularly in terms of its photosynthetic membrane system and in its light absorption properties.

Seasonal patterns of nitrogen fixation and denitrification in oceanic mangrove habitats

Mangrove peat soils are home to a variety of microbial communities that may play a vital role in system-level elemental cycling. We examined rates of nitrogen fixation and denitrification in benthic microbial mats on Twin Cays, Belize, a pair of oceanic mangrove islands. A tree-height gradient across the islands created distinct habitats for benthic microbes. Seawater flushing of the benthos and tree height decreased landward from tall, dense trees on the island fringe through a transition zone of high elevation and intermediate tree heights. In the center of the islands, microbial mats with dense communities of cyanobacteria and purple sulfur bacteria covered the benthic surface of shallow ponds and around dwarf trees. Wet-dry seasonality, tidal cycles and elevation controlled the extent of mat exposure to desiccation and UV radiation. Nitrogen fixation was controlled primar- ily by the sensitivity of nitrogenase to oxygen inhibition, whereas denitrification was limited by oxi- dant (nitrate) availability. Diel patterns of nitrogen fixation varied with the type of cyanobacteria dominant in each mat. Dissolved inorganic nitrogen concentration influenced both nitrogen fixation and denitrification rates. Redox conditions contributed to variability in mat nitrogen fixation and denitrification response to nutrient addition, while dissolved organic carbon did not. Microbial mat nitrogen cycling likely contributes to the nutrient (nitrogen and phosphorus) limitation patterns observed in the mangrove trees; in dwarf habitats, mats serve as a source of nitrogen via nitrogen fixation, while in fringe and transition habitats, mats compete with the trees for nitrogen via denitrification.