Rhodoferax Ferrireducens Research Articles

Rhodoferax lithotrophicus sp. nov., a neutrophilic iron-oxidizing and -reducing bacterium isolated from iron-rich freshwater sediments.

A neutrophilic iron-oxidizing and -reducing bacterium, strain MIZ03T, was previously isolated from a wetland in Ibaraki, Japan. Here, we report the detailed characteristics of this strain. It was motile with a single polar flagellum, and Gram-stain-negative. It could grow not only chemolithoautotrophically but also chemoorganotrophically by aerobic respiration and fermentation. Major cellular fatty acids were C16 : 1 ω7c/C16 : 1 ω6c, and C16 : 0. Phylogenetic analyses indicated that strain MIZ03T belonged to the genus Rhodoferax. This strain was closely related to Rhodoferax ferrireducens with 98.5 % of 16S rRNA gene sequence similarity. Based on its phenotypic and genomic based characteristics, we conclude that strain MIZ03T represents a new species in the genus Rhodoferax. We propose the name Rhodoferax lithotrophicus sp. nov. to accommodate this strain. The type strain is MIZ03T (=JCM 34246T=DSM 113266T). We also propose the name Rhodoferax koreensis sp. nov., of which the type strain is DCY110T (=KCTC 52288T=JCM 31441T), for the effectively, but not yet validly, published name 'Rhodoferax koreense'.

GEM-based computational modeling for exploring metabolic interactions in a microbial community.

Microbial communities play fundamental roles in every complex ecosystem, such as soil, sea and the human body. The stability and diversity of the microbial community depend precisely on the composition of the microbiota. Any change in the composition of these communities affects microbial functions. An important goal of studying the interactions between species is to understand the behavior of microbes and their responses to perturbations. These interactions among species are mediated by the exchange of metabolites within microbial communities. We developed a computational model for the microbial community that has a separate compartment for exchanging metabolites. This model can predict possible metabolites that cause competition, commensalism, and mutual interactions between species within a microbial community. Our constraint-based community metabolic modeling approach provides insights to elucidate the pattern of metabolic interactions for each common metabolite between two microbes. To validate our approach, we used a toy model and a syntrophic co-culture of Desulfovibrio vulgaris and Methanococcus maripaludis, as well as another in co-culture between Geobacter sulfurreducens and Rhodoferax ferrireducens. For a more general evaluation, we applied our algorithm to the honeybee gut microbiome, composed of seven species, and the epiphyte strain Pantoea eucalypti 299R. The epiphyte strain Pe299R has been previously studied and cultured with six different phyllosphere bacteria. Our algorithm successfully predicts metabolites, which imply mutualistic, competitive, or commensal interactions. In contrast to OptCom, MRO, and MICOM algorithms, our COMMA algorithm shows that the potential for competitive interactions between an epiphytic species and Pe299R is not significant. These results are consistent with the experimental measurements of population density and reproductive success of the Pe299R strain.

Reaction pathways and Sb(III) minerals formation during the reduction of Sb(V) by Rhodoferax ferrireducens strain YZ-1

Antimony (Sb), a non-essential metalloid, can be released into the environment through various industrial activities. Sb(III) is considered more toxic than Sb(V), but Sb(III) can be immobilized through the precipitation of insoluble Sb2S3 or Sb2O3. In the subsurface, Sb redox chemistry is largely controlled by microorganisms; however, the exact mechanisms of Sb(V) reduction to Sb(III) are still unclear. In this study, a new strain of Sb(V)-reducing bacterium, designated as strain YZ-1, that can respire Sb(V) as a terminal electron acceptor was isolated from Sb-contaminated soils. 16S-rRNA gene sequencing of YZ-1 revealed high similarity to a known Fe(III)-reducer, Rhodoferax ferrireducens. XRD and XAFS analyses revealed that bioreduction of Sb(V) to Sb(III) proceed through a transition from amorphous valentinite to crystalline senarmontite (allotropes of Sb2O3). Genomic DNA sequencing found that YZ-1 possesses arsenic (As) metabolism genes, including As(V) reductase arsC. The qPCR analysis showed that arsC was highly expressed during Sb(V)-reduction by YZ-1, and thus is proposed as the potential Sb(V) reductase in YZ-1. This study provides new insight into the pathways and products of microbial Sb(V) reduction and demonstrates the potential of a newly isolated bacterium for Sb bioremediation.

Overexpression of Dioxygenase Encoding Gene Accelerates the Phenolic Aldehyde Conversion and Ethanol Fermentability of Zymomonas mobilis.

NADH-dependent reductase enzyme catalyzes the phenolic aldehyde conversion and correspondingly improves the ethanol fermentability of the ethanologenic Zymomonas mobilis. This study constructed the transcriptional landscape of mono/dioxygenase genes in Z. mobilis ZM4 under the stress of the toxic phenolic aldehyde inhibitors of 4-hydroxybenzaldehyde, syringaldehyde, and vanillin. One specific dioxygenase encoding gene ZMO1721 was differentially expressed by 3.07-folds under the stress of 4-hydroxybenzaldehyde among the eleven mono/dioxygenase genes. The purified ZMO1721 shared 99.9% confidence and 48.0% identity with the oxidoreductase in Rhodoferax ferrireducens T118 was assayed and the NADH-dependent reduction activity was confirmed for phenolic aldehyde vanillin conversion. The ZMO1721 gene was then overexpressed in Z. mobilis ZM4 and the 4-hydroxybenzaldehyde conversion rate was accelerated. The cell growth, glucose consumption, and ethanol productivity of Z. mobilis ZM4 were also improved by ZMO1721 overexpression. The genes identified on improving phenolic aldehyde tolerance and ethanol fermentability in this study could be used as the synthetic biology tools for modification of ethanologenic strains.

Lithogenic hydrogen supports microbial primary production in subglacial and proglacial environments

Life in environments devoid of photosynthesis, such as on early Earth or in contemporary dark subsurface ecosystems, is supported by chemical energy. How, when, and where chemical nutrients released from the geosphere fuel chemosynthetic biospheres is fundamental to understanding the distribution and diversity of life, both today and in the geologic past. Hydrogen (H2) is a potent reductant that can be generated when water interacts with reactive components of mineral surfaces such as silicate radicals and ferrous iron. Such reactive mineral surfaces are continually generated by physical comminution of bedrock by glaciers. Here, we show that dissolved H2 concentrations in meltwaters from an iron and silicate mineral-rich basaltic glacial catchment were an order of magnitude higher than those from a carbonate-dominated catchment. Consistent with higher H2 abundance, sediment microbial communities from the basaltic catchment exhibited significantly shorter lag times and faster rates of net H2 oxidation and dark carbon dioxide (CO2) fixation than those from the carbonate catchment, indicating adaptation to use H2 as a reductant in basaltic catchments. An enrichment culture of basaltic sediments provided with H2, CO2, and ferric iron produced a chemolithoautotrophic population related to Rhodoferax ferrireducens with a metabolism previously thought to be restricted to (hyper)thermophiles and acidophiles. These findings point to the importance of physical and chemical weathering processes in generating nutrients that support chemosynthetic primary production. Furthermore, they show that differences in bedrock mineral composition can influence the supplies of nutrients like H2 and, in turn, the diversity, abundance, and activity of microbial inhabitants.

A Novel "Microbial Bait" Technique for Capturing Fe(III)-Reducing Bacteria.

Microbial reduction of Fe(III) is a key geochemical process in anoxic environments, controlling the degradation of organics and the mobility of metals and radionuclides. To further understand these processes, it is vital to develop a reliable means of capturing Fe(III)-reducing microorganisms from the field for analysis and lab-based investigations. In this study, a novel method of capturing Fe(III)-reducing bacteria using Fe(III)-coated pumice “microbe-baits” was demonstrated. The methodology involved the coating of pumice (approximately diameter 4 to 6 mm) with a bioavailable Fe(III) mineral (akaganeite), and was verified by deployment into a freshwater spring for 2 months. On retrieval, the coated pumice baits were incubated in a series of lab-based microcosms, amended with and without electron donors (lactate and acetate), and incubated at 20°C for 8 weeks. 16S rRNA gene sequencing using the Illumina MiSeq platform showed that the Fe(III)-coated pumice baits, when incubated in the presence of lactate and acetate, enriched for Deltaproteobacteria (relative abundance of 52% of the sequences detected corresponded to Geobacter species and 24% to Desulfovibrio species). In the absence of added electron donors, Betaproteobacteria were the most abundant class detected, most heavily represented by a close relative to Rhodoferax ferrireducens (15% of species detected), that most likely used organic matter sequestered from the spring waters to support Fe(III) reduction. In addition, TEM-EDS analysis of the Fe(III)-coated pumice slurries amended with electron donors revealed that a biogenic Fe(II) mineral, magnetite, was formed at the end of the incubation period. These results demonstrate that Fe(III)-coated pumice “microbe baits” can potentially help target metal-reducing bacteria for culture-dependent studies, to further our understanding of the nano-scale microbe-mineral interactions in aquifers.

Microbial reduction of Fe(III) coupled to the biodegradation of isosaccharinic acid (ISA)

Isosaccharinic acid (ISA) forms through alkaline hydrolysis of cellulose in intermediate level nuclear waste and is a strong complexant of metals with the potential to mobilize priority radionuclides in an underground geological disposal facility (GDF). In this study, microbial ISA degradation was studied under anaerobic conditions with Fe(III) oxyhydroxide as the terminal electron acceptor at pH 7 to 10, representative of conditions in the geosphere surrounding a geological disposal facility. A multidisciplinary approach was used to study the fate and limits of ISA biodegradation, including mineralogical analyses (XRD, ESEM and TEM), alongside geochemical profiling of ISA and its biodegradation products, and microbial community analysis by 16S rRNA gene sequencing. Under the conditions imposed, ISA degradation was constrained to a pH limit of ≤9 and was degraded via Fe(III) reduction and fermentation. Biominerals, resulting from ISA degradation, were analyzed. These revealed two Fe(II) minerals, siderite [FeCO3] and vivianite [(Fe3PO4)2·8H2O], not normally associated with the microbial reduction of ferrihydrite in laboratory cultures, possibly as a result of chelation of ISA with Fe(III) that made the electron acceptor more soluble. After the enrichment at pH 7, a complex microbial consortium, including Clostridia and close relatives to known Fe(III)-reducing bacteria Geobacter sp. and Rhodoferax ferrireducens, was associated with these activities. The results are discussed in the context of geological disposal safety case development, including the potential impact on priority radionuclides including U(VI), Np(V), and Tc(VII), which are highly susceptible to reductive immobilization when in contact with Fe(II)-bearing biominerals.

Identification and Characterization of a Dominant Sulfolane-Degrading Rhodoferax sp. via Stable Isotope Probing Combined with Metagenomics

Sulfolane is an industrial solvent and emerging organic contaminant affecting groundwater around the world, but little is known about microbes capable of biodegrading sulfolane or the pathways involved. We combined DNA-based stable isotope probing (SIP) with genome-resolved metagenomics to identify microorganisms associated with sulfolane biodegradation in a contaminated subarctic aquifer. In addition to 16S rRNA gene amplicon sequencing, we performed shotgun metagenomics on the 13C-labeled DNA to obtain functional and taxonomic information about the active sulfolane-degrading community. We identified the primary sulfolane degrader, comprising ~85% of the labeled community in the amplicon sequencing dataset, as closely related to Rhodoferax ferrireducens strain T118. We obtained a 99.8%-complete metagenome-assembled genome for this strain, allowing us to identify putative pathways of sulfolane biodegradation. Although the 4S dibenzothiophene desulfurization pathway has been proposed as an analog for sulfolane biodegradation, we found only a subset of the required genes, suggesting a novel pathway specific to sulfolane. DszA, the enzyme likely responsible for opening the sulfolane ring structure, was encoded on both the chromosome and a plasmid. This study demonstrates the power of integrating DNA-SIP with metagenomics to characterize emerging organic contaminant degraders without culture bias and expands the known taxonomic distribution of sulfolane biodegradation.

Rhodoferax koreense sp. nov, an obligately aerobic bacterium within the family Comamonadaceae, and emended description of the genus Rhodoferax.

Gram-staining-negative, uniflagellated, rod-shaped, designated as DCY110T, was isolated from sludge located in Gangwon province, Republic of Korea. The phylogenetic tree of 16S rRNA gene sequence showed that the strain DCY110T belonged to the genus Rhodoferax with a close similarity to Rhodoferax saidenbachensis DSM 22694T (97.7%), Rhodoferax antarcticus DSM 24876T (97.5%), Rhodoferax ferrireducens DSM 15236T (97.3%), and Rhodoferax fermentans JCM 7819T (96.7%). The predominant isoprenoid quinine was ubiquinone (Q-8). DNA G + C content was 62.8 mol%. The major polar lipids were phosphatidylethanolamine and two unidentified phospholipids. The major fatty acids (> 10%) were C12:0, C16:0, summed feature 3 (which comprised C16:1 ω7c and/or C16:1 ω6c). The DNA-DNA relatedness values between the strain DCY110T and the closely related relatives used in this study were lower than 70%. Based on the following polyphasic analysis, the strain DCY110T is considered as a novel species of the genus Rhodoferax, for which the name Rhodoferax koreense sp. nov. is proposed. The type strain is DCY-110T (= KCTC 52288T = JCM 31441T).

Harvesting Energy using Biocatalysts

<i>Harvesting Energy using Biocatalysts</i>

D-OptCom: Dynamic multi-level and multi-objective metabolic modeling of microbial communities.

Most microbial communities change with time in response to changes and/or perturbations in environmental conditions. Temporal variations in interspecies metabolic interactions within these communities can significantly affect their structure and function. Here, we introduce d-OptCom, an extension of the OptCom procedure, for the dynamic metabolic modeling of microbial communities. It enables capturing the temporal dynamics of biomass concentration of the community members and extracellular concentration of the shared metabolites, while integrating species- and community-level fitness functions. The applicability of d-OptCom was demonstrated by modeling the dynamic co-growth of auxotrophic mutant pairs of E. coli and by computationally assessing the dynamics and composition of a uranium-reducing community comprised of Geobacter sulfurreducens, Rhodoferax ferrireducens, and Shewanella oneidensis. d-OptCom was also employed to examine the impact of lactate vs acetate addition on the relative abundance of uranium-reducing species. These studies highlight the importance of simultaneously accounting for both species- and community-level fitness functions when modeling microbial communities, and demonstrate that the incorporation of uptake kinetic information can substantially improve the prediction of interspecies flux trafficking. Overall, this study paves the way for the dynamic multi-level and multi-objective analysis of microbial ecosystems.

Rhodoferax saidenbachensis sp. nov., a psychrotolerant, very slowly growing bacterium within the family Comamonadaceae, proposal of appropriate taxonomic position of Albidiferax ferrireducens strain T118T in the genus Rhodoferax and emended description of the genus Rhodoferax

A Gram-stain-negative, oxidase and phosphatase-positive and catalase-negative, short rod-shaped bacterium was isolated from sediment of a drinking water reservoir in Germany. Based on 16S rRNA gene sequence and phenotypic properties, the bacterium belongs to the genus Rhodoferax within the family Comamonadaceae. The new taxon differed from related species mainly with respect to its fatty acid composition, low growth temperature, lack of pigments in young cultures and ability to utilize glycerol and d-mannose but not urea. The major fatty acids were C16 : 1ω7c and/or iso-C15 : 0 2-OH, C16 : 0, and C18 : 1ω7c. The only ubiquinone detected was ubiquinone Q-8. The DNA G+C content was 60.3-61 mol%. Because of the phenotypic and genotypic differences from the most closely related taxa, the new strain represents a novel species for which the name Rhodoferax saidenbachensis sp. nov. is proposed. The type strain is ED16(T) ( = CCUG 57711(T) = ATCC BAA-1852(T) = DSM 22694(T)). An emended description of the genus Rhodoferax is proposed. Based on the results of this study, strain T118(T) (Albidiferax ferrireducens) is properly placed in the genus Rhodoferax as Rhodoferax ferrireducens.

분자생물학적 방법을 이용한 지열시스템 관정 및 주변지역 미생물종 모니터링

This study was conducted to monitor microbial species dynamics within the aquifer due to long term operation of geothermal heat pump system. The species were identified by molecular biological methods of 16S rDNA. Groundwater sample was collected from both open (S region) and closed geothermal recovery system (J region) along with the control. J measured and control as well as S measured found Ralstonia pickettii as dominant species at year 2010. In contrast, Rhodoferax ferrireducens was dominantly observed for the control of S. In 2011, Sediminibacterium sp. was universely identified as the dominant species regardless of the monitoring places and type of sample, i.e., measured or control. The difference in the dynamics between the measured and the control was not critically observed, but annual variation was more strikingly found. It reveals that possible environmental changes (e.g. ORP and DO) due to the operation of geothermal heat recovery system in aquifer could be more exceedingly preceded to differentiate annual variation of microbial species rather than positional differences.

Mtr extracellular electron-transfer pathways in Fe(III)-reducing or Fe(II)-oxidizing bacteria: a genomic perspective

Originally discovered in the dissimilatory metal-reducing bacterium Shewanella oneidensis MR-1 (MR-1), key components of the Mtr (i.e. metal-reducing) pathway exist in all strains of metal-reducing Shewanella characterized. The protein components identified to date for the Mtr pathway of MR-1 include four multihaem c-Cyts (c-type cytochromes), CymA, MtrA, MtrC and OmcA, and a porin-like outer membrane protein MtrB. They are strategically positioned along the width of the MR-1 cell envelope to mediate electron transfer from the quinone/quinol pool in the inner membrane to Fe(III)-containing minerals external to the bacterial cells. A survey of microbial genomes has identified homologues of the Mtr pathway in other dissimilatory Fe(III)-reducing bacteria, including Aeromonas hydrophila, Ferrimonas balearica and Rhodoferax ferrireducens, and in the Fe(II)-oxidizing bacteria Dechloromonas aromatica RCB, Gallionella capsiferriformans ES-2 and Sideroxydans lithotrophicus ES-1. The apparent widespread distribution of Mtr pathways in both Fe(III)-reducing and Fe(II)-oxidizing bacteria suggests a bidirectional electron transfer role, and emphasizes the importance of this type of extracellular electron-transfer pathway in microbial redox transformation of iron. The organizational and electron-transfer characteristics of the Mtr pathways may be shared by other pathways used by micro-organisms for exchanging electrons with their extracellular environments.

Enrichment and characterization of a bacteria consortium capable of heterotrophic nitrification and aerobic denitrification at low temperature

Nitrogen removal in wastewater treatment plants is usually severely inhibited under cold temperature. The present study proposes bioaugmentation using psychrotolerant heterotrophic nitrification–aerobic denitrification consortium to enhance nitrogen removal at low temperature. A functional consortium has been successfully enriched by stepped increase in DO concentration. Using this consortium, the specific removal rates of ammonia and nitrate at 10°C reached as high as 3.1mgN/(gSSh) and 9.6mgN/(gSSh), respectively. PCR-DGGE and clone library analysis both indicated a significant reduction in bacterial diversity during enrichment. Phylogenetic analysis based on nearly full-length 16S rRNA genes showed that Alphaproteobacteria, Deltaproteobacteria and particularly Bacteroidetes declined while Gammaproteobacteria (all clustered into Pseudomonas sp.) and Betaproteobacteria (mainly Rhodoferax ferrireducens) became dominant in the enriched consortium. It is likely that Pseudomonas spp. played a major role in nitrification and denitrification, while R. ferrireducens and its relatives utilized nitrate as both electron acceptor and nitrogen source.

Characterization and transcription of arsenic respiration and resistance genes during in situ uranium bioremediation.

The possibility of arsenic release and the potential role of Geobacter in arsenic biogeochemistry during in situ uranium bioremediation was investigated because increased availability of organic matter has been associated with substantial releases of arsenic in other subsurface environments. In a field experiment conducted at the Rifle, CO study site, groundwater arsenic concentrations increased when acetate was added. The number of transcripts from arrA, which codes for the α-subunit of dissimilatory As(V) reductase, and acr3, which codes for the arsenic pump protein Acr3, were determined with quantitative reverse transcription-PCR. Most of the arrA (>60%) and acr3-1 (>90%) sequences that were recovered were most similar to Geobacter species, while the majority of acr3-2 (>50%) sequences were most closely related to Rhodoferax ferrireducens. Analysis of transcript abundance demonstrated that transcription of acr3-1 by the subsurface Geobacter community was correlated with arsenic concentrations in the groundwater. In contrast, Geobacter arrA transcript numbers lagged behind the major arsenic release and remained high even after arsenic concentrations declined. This suggested that factors other than As(V) availability regulated the transcription of arrA in situ, even though the presence of As(V) increased the transcription of arrA in cultures of Geobacter lovleyi, which was capable of As(V) reduction. These results demonstrate that subsurface Geobacter species can tightly regulate their physiological response to changes in groundwater arsenic concentrations. The transcriptomic approach developed here should be useful for the study of a diversity of other environments in which Geobacter species are considered to have an important influence on arsenic biogeochemistry.

Molecular analysis of spatial variation of iron-reducing bacteria in riverine alluvial aquifers of the Mankyeong River

Alluvial aquifers are one of the mainwater resources in many countries. Iron reduction in alluvial aquifers is often a major anaerobic process involved in bioremediation or causing problems, including the release of As trapped in Fe(III) oxide. We investigated the distribution of potential iron-reducing bacteria (IRB) in riverine alluvial aquifers (B1, B3, and B6 sites) at the Mankyeong River, Republic of Korea. Inactive iron reduction zones, the diversity and abundance of IRB can be examined using a clone library and quantitative PCR analysis of 16S rRNA genes. Geobacter spp. are potential IRB in the iron-reducing zone at the B6 (9 m) site, where high Fe(II) and arsenic (As) concentrations were observed. At the B3 (16 m) site, where low iron reduction activity was predicted, a dominant clone (10.6%) was 99% identical in 16S rRNA gene sequence with Rhodoferax ferrireducens. Although a major clone belonging to Clostridium spp. was found, possible IRB candidates could not be unambiguously determined at the B1 (18 m) site. Acanonical correspondence analysis demonstrated that, among potential IRB, only the Geobacteraceae were well correlated with Fe(II) and As concentrations. Our results indicate high environmental heterogeneity, and thus high spatial variability, in thedistribution of potential IRB in the riverine alluvial aquifersnear the Mankyeong River.

Microbial and geochemical features suggest iron redox cycling within bacteriogenic iron oxide-rich sediments

The geochemistry, mineralogy and microbiology of bacteriogenic iron oxides (BIOS) deposited at a circumneutral groundwater seep were examined to evaluate the biogeochemical controls on the redox cycle of iron in such environments. X-ray diffraction and extended X-ray absorption fine structure (EXAFS) spectroscopy showed that 2-line ferrihydrite dominated (60–70%) the BIOS mineralogy, with more crystalline lepidocrocite and goethite phases comprising the remainder, likely the product of Fe(II)-induced dissolution–reprecipitation pathways. Subsurface porewater profiles displayed an increase in dissolved (<0.2μm) equilibrium concentrations of Fe(II), also reflected in peaks in HCl-extractable sedimentary Fe(II) at depth, indicative of biological iron reduction. Interestingly EXAFS analysis of sediments collected from such depths revealed little mineralogical change from the surficial BIOS. Dissolved concentrations of Fe(III), likely present as nanoparticulates or organic complexes, also rose with depth. Scanning electron microscopy images of the surface BIOS samples showed a proliferation of sheath structures characteristic of Fe(II)-oxidizing Leptothrix spp. A 16S rRNA clone library obtained from the surficial BIOS included microorganisms closely associated to known Fe(II)-oxidizing (Gallionella spp., Sideroxydans spp.) and Fe(III)-reducing (Rhodoferax ferrireducens and Geothrix fermentans) bacteria. Analysis of subsurface sediment collected from an Fe(III)-reducing horizon revealed a drop in the proportion of β-Proteobacteria, which hosts numerous Fe(II)-oxidizing genera, and a rise in the δ-Proteobacteria, home to anaerobic metal-reducing prokaryotes. The recognition of clones with close identity to bacteria intimately involved in complementary iron redox transformations in the same surficial BIOS sediment suggests that microbial oxidation and reduction of iron may be tightly coupled in this, and other, shallow sedimentary environments.

Reduction of iron (III) and humic substances plays a major role in anaerobic respiration in an Arctic peat soil

Arctic peat soils contain vast reserves of organic C and are largely anaerobic. However, anaerobic respiration, particularly the role of Fe(III) and humic substances as electron acceptors, is not well understood in such ecosystems. We investigated these processes in a drained thaw lake basin on the Arctic coastal plain near Barrow, Alaska. We measured concentrations of soluble Fe and other potential electron acceptors, described the microbial community, and performed experiments in the laboratory and field to measure net rates of Fe(III) reduction and the relationship of this process to C cycling. In most areas within the basin, aerobic conditions existed only in the upper few centimeters of soil, though oxygen penetrated deeper in raised areas, such as rims of ice wedge polygons. Concentrations of nitrate and sulfate in soil pore water were low or negligible. Soil pore water contained surprisingly high concentrations of Fe(II) and Fe(III), in the range of hundreds of μM, suggesting the presence of organic chelators. The solid phase contained substantial amounts of iron minerals, with a progressively reduced oxidation state throughout the growing season. The most abundant 16S rRNA sequence in our gene survey was closely related to the Fe(III)‐reducing bacterium, Rhodoferax ferrireducens, and other sequences closely related to Fe‐transforming bacteria were found. Field and laboratory incubations with soluble Fe(III) and the quinonic compound, AQDS (a common humic analog), stimulated respiration and verified that Fe(III) reduction occurs in these soils. We conclude that reduction of Fe(III) and humic substances are major metabolic pathways in this ecosystem.

Genome-scale dynamic modeling of the competition between Rhodoferax and Geobacter in anoxic subsurface environments

The advent of rapid complete genome sequencing, and the potential to capture this information in genome-scale metabolic models, provide the possibility of comprehensively modeling microbial community interactions. For example, Rhodoferax and Geobacter species are acetate-oxidizing Fe(III)-reducers that compete in anoxic subsurface environments and this competition may have an influence on the in situ bioremediation of uranium-contaminated groundwater. Therefore, genome-scale models of Geobacter sulfurreducens and Rhodoferax ferrireducens were used to evaluate how Geobacter and Rhodoferax species might compete under diverse conditions found in a uranium-contaminated aquifer in Rifle, CO. The model predicted that at the low rates of acetate flux expected under natural conditions at the site, Rhodoferax will outcompete Geobacter as long as sufficient ammonium is available. The model also predicted that when high concentrations of acetate are added during in situ bioremediation, Geobacter species would predominate, consistent with field-scale observations. This can be attributed to the higher expected growth yields of Rhodoferax and the ability of Geobacter to fix nitrogen. The modeling predicted relative proportions of Geobacter and Rhodoferax in geochemically distinct zones of the Rifle site that were comparable to those that were previously documented with molecular techniques. The model also predicted that under nitrogen fixation, higher carbon and electron fluxes would be diverted toward respiration rather than biomass formation in Geobacter, providing a potential explanation for enhanced in situ U(VI) reduction in low-ammonium zones. These results show that genome-scale modeling can be a useful tool for predicting microbial interactions in subsurface environments and shows promise for designing bioremediation strategies.