Autotrophic Pathways Research Articles

Intertwining of the C-N-S cycle in passive and aerated constructed wetlands.

The microbial processes occurring in constructed wetlands (CWs) are difficult to understand owing to the complex interactions occurring between a variety of substrates, microorganisms, and plants under the given physicochemical conditions. This frequently leads to very large unexplained nitrogen losses in these systems. In continuation of our findings on Anammox contributions, our research on full-scale field CWs has suggested the significant involvement of the sulfur cycle in the conventional C-N cycle occurring in wetlands, which might closely explain the nitrogen losses in these systems. This paper explored the possibility of the sulfur-driven autotrophic denitrification (SDAD) pathway in different types of CWs, shallow and deep and passive and aerated systems, by analyzing the metagenomic bacterial communities present within these CWs. The results indicate a higher abundance of SDAD bacteria (Paracoccus and Arcobacter) in deep passive systems compared to shallow systems and presence of a large number of SDAD genera (Paracoccus, Thiobacillus, Beggiatoa, Sulfurimonas, Arcobacter, and Sulfuricurvum) in aerated CWs. The bacteria belonging to the functional category of dark oxidation of sulfur compounds were found to be enriched in deep and aerated CWs hinting at the possible role of the SDAD pathway in total nitrogen removal in these systems. As a case study, the percentage nitrogen removal through SDAD pathway was calculated to be 15-20% in aerated wetlands. The presence of autotrophic pathways for nitrogen removal can prove highly beneficial in terms of reducing sludge generation and hence reducing clogging, making aerated CWs a sustainable wastewater treatment solution.

Metabolic adaptations underpin high productivity rates in relict subsurface water

Groundwater aquifers are ecological hotspots with diverse microbes essential for biogeochemical cycles. Their ecophysiology has seldom been studied on a basin scale. In particular, our knowledge of chemosynthesis in the deep aquifers where temperatures reach 60 °C, is limited. Here, we investigated the diversity, activity, and metabolic potential of microbial communities from nine wells reaching ancient groundwater beneath Israel’s Negev Desert, spanning two significant, deep (up to 1.5 km) aquifers, the Judea Group carbonate and Kurnub Group Nubian sandstone that contain fresh to brackish, hypoxic to anoxic water. We estimated chemosynthetic productivity rates ranging from 0.55 ± 0.06 to 0.82 ± 0.07 µg C L−1 d−1 (mean ± SD), suggesting that aquifer productivity may be underestimated. We showed that 60% of MAGs harbored genes for autotrophic pathways, mainly the Calvin–Benson–Bassham cycle and the Wood–Ljungdahl pathway, indicating a substantial chemosynthetic capacity within these microbial communities. We emphasize the potential metabolic versatility in the deep subsurface, enabling efficient carbon and energy use. This study set a precedent for global aquifer exploration, like the Nubian Sandstone Aquifer System in the Arabian and Western Deserts, and reconsiders their role as carbon sinks.

Description of Fuscovulum ytuae sp. nov, a facultative autotroph isolated from the intertidalite of Yangma island, China.

In this study, we reported a Gram-stain-negative, ovoid to rod-shaped, atrichous, and facultative anaerobe bacteria strain named YMD61T, which was isolated from the intertidal sediment of Yangma island, China. Growth of strain YMD61T occurred at 10.0-45.0°C (optimum, 30.0°C), pH 7.0-10.0 (optimum, 8.0) and with 0-3.0% (w/v) NaCl (optimum, 2.0%). Phylogenetic tree analysis based on 16S rRNA gene or genomic sequence indicated that strain YMD61T belonged to the genus Fuscovulum and was closely related to Fuscovulum blasticum ATCC 33,485T (96.6% sequence similarity). Genomic analysis indicated that strain YMD61T contains a circular chromosome of 3,895,730bp with DNA G + C content of 63.3%. The genomic functional analysis indicated that strain YMD61T is a novel sulfur-metabolizing bacteria, which is capable of fixing carbon through an autotrophic pathway by integrating the processes of photosynthesis and sulfur oxidation. The predominant respiratory quinone of YMD61T was ubiquinone-10 (Q-10). The polar lipids of YMD61T contained phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, five unidentified lipids, unidentified aminolipid and unidentified aminophospholipid. The major fatty acids of strain YMD61T contained C18:1ω7c 11-methyl and summed feature 8 (C18:1 ω 7c or/and C18:1 ω 6c). Phylogenetic, physiological, biochemical and morphological analyses suggested that strain YMD61T represents a novel species of the genus Fuscovulum, and the name Fuscovulum ytuae sp. nov. is proposed. The type strain is YMD61T (= MCCC 1K08483T = KCTC 43,537T).

Widespread dissolved inorganic carbon-modifying toolkits in genomes of autotrophic Bacteria and Archaea and how they are likely to bridge supply from the environment to demand by autotrophic pathways.

Using dissolved inorganic carbon (DIC) as a major carbon source, as autotrophs do, is complicated by the bedeviling nature of this substance. Autotrophs using the Calvin-Benson-Bassham cycle (CBB) are known to make use of a toolkit comprised of DIC transporters and carbonic anhydrase enzymes (CA) to facilitate DIC fixation. This minireview provides a brief overview of the current understanding of how toolkit function facilitates DIC fixation in Cyanobacteria and some Proteobacteria using the CBB and continues with a survey of the DIC toolkit gene presence in organisms using different versions of the CBB and other autotrophic pathways (reductive citric acid cycle, Wood-Ljungdahl pathway, hydroxypropionate bicycle, hydroxypropionate-hydroxybutyrate cycle, and dicarboxylate-hydroxybutyrate cycle). The potential function of toolkit gene products in these organisms is discussed in terms of CO2 and HCO3- supply from the environment and demand by the autotrophic pathway. The presence of DIC toolkit genes in autotrophic organisms beyond those using the CBB suggests the relevance of DIC metabolism to these organisms and provides a basis for better engineering of these organisms for industrial and agricultural purposes.

Flow event size influences carbon, nutrient and zooplankton dynamics in a highly regulated lowland river

AbstractRiver regulation and water extraction has significantly altered flow regimes and reduced flood events in many inland river systems. Environmental flows have been adopted in many systems to mitigate the ecological impacts of river regulation, however a lack of knowledge regarding the interrelationship between flow regimes, carbon transport and instream productivity make prioritising water management difficult. To address this knowledge gap, we conducted a study on the Namoi River in the Murray-Darling Basin, Australia, monitoring changes in dissolved organic carbon (DOC), nutrient dynamics and planktonic food web structure during a period of variable flows. Nutrient and DOC concentrations were positively correlated with river discharge and zooplankton concentrations were highest post flow events. Planktonic chlorophyll-a, increased DOC concentration and higher discharge were the most influential drivers of change in zooplankton communities. Further, our results indicated that flow events increased production through both heterotrophic and autotrophic pathways, significantly boosting zooplankton concentration compared to base flow conditions across all measured flow events. We suggest even small in-channel flow events can be important for increasing basal and zooplankton production in rivers, and therefore should be protected or promoted by environmental flow management, particularly during drought conditions.

Nitrogen removal by algal-bacterial consortium during mainstream wastewater treatment: Transformation mechanisms and potential N2O mitigation

This work investigated nitrogen transformation pathways of the algal-bacterial consortium as well as its potential in reducing nitrous oxide (N2O) emission in enclosed, open and aerated reactors. The results confirmed the superior ammonium removal performance of the algal-bacterial consortium relative to the single algae (Chlorella vulgaris) or the activated sludge, achieving the highest efficiency at 100% and the highest rate of 7.34 mg N g MLSS−1 h−1 in the open reactor with glucose. Enhanced total nitrogen (TN) removal (to 74.6%) by the algal-bacterial consortium was achieved via mixotrophic algal assimilation and bacterial denitrification under oxygen-limited and glucose-sufficient conditions. Nitrogen distribution indicated that ammonia oxidation (∼41.8%) and algal assimilation (∼43.5%) were the main pathways to remove ammonium by the algal-bacterial consortium. TN removal by the algal-bacterial consortium was primarily achieved by algal assimilation (28.1–40.8%), followed by bacterial denitrification (2.9–26.5%). Furthermore, the algal-bacterial consortium contributed to N2O mitigation compared with the activated sludge, reducing N2O production by 35.5–55.0% via autotrophic pathways and by 81.0–93.6% via mixotrophic pathways. Nitrogen assimilation by algae was boosted with the addition of glucose and thus largely restrained N2O production from nitrification and denitrification. The synergism between algae and bacteria was also conducive to an enhanced N2O reduction by denitrification and reduced direct/indirect carbon emissions.

Charcoal stabilizes pH and improves nutrients removal of bacteria-microalgae interaction: A potential for improving water quality in aquaculture

Aquaculture is prone to pH fluctuations depending on dominating microbial pathway in the management. The present study takes advantage of the influence of pH in changing the properties of charcoal to condition the pH and improve nutrient removal. Two Synthetic aquaculture wastewaters of 100 mg/L TAN were designed to test the pathway's nutrient removal and pH stability. Glucose at 15 C/N was added to one wastewater, favoring heterotrophic pathway. The other was free of organic carbon, favoring autotrophic pathway. Powdered charcoal was added to each pathway at 0.5, 1.0, 2.0, and 4.0% (charcoal weight/volume of media) compared to the 0 % group without charcoal. The result revealed that as charcoal percentages increase, the pH shifts towards stabilization and improves nutrient removal (<i>p</i> ≤ 0.05). At 4.0% charcoal, the pH was stabilized at 6.0 – 7.5 in heterotrophic and 7.5 – 7.9 in autotrophic pathways, which are within the optimum range for the survival of aquatic organisms, corresponding to the removal efficiency of TAN (99.96 and 50 %), and phosphate (55 and 17 %) within 6 days. Hence, for every 100 mg/L of TAN in wastewater, 4.0% of the wastewater volume as charcoal could stabilize pH and improve microbial nutrient removal.

Insights on Microbial Communities Inhabiting Non-Volcanic Hot Springs.

The northwest of Spain has an abundance of non-volcanic hot springs that, until recently, had only been used for thermalism activities. One of such hot springs, Muiño da Veiga, has now been explored using metagenomics to study the microbial community that inhabits these high-temperature circumneutral continental waters. Sequencing of the metagenome allowed the characterization of its composition, diversity, metabolic connections and potential as a source for thermozymes, as well as its ability to assemble MAGs. A diverse microbial community dominated by Bacteria domain members was revealed, particularly from the early-branching Aquificales group. The most abundant genus was Sulfurihydrogenibium, known for its implication in sulfur cycling and for forming mats that enable novel niches. The variety of primary producers with autotrophic pathways (and specifically the sulfur oxidizing pathway) expands the range of available nutrients, and the increase in biomass forms thicker mats, resulting in more available niches and broader microbial diversity. Nonetheless, certain metabolic pathways were attributed to less abundant members of the microbial community, reinforcing the idea that the rare biosphere plays important roles in the network of interactions present in an ecosystem and acts as genetic reservoirs. In addition, three of the assembled MAGs represent novel microbial diversity found in this hot spring. Moreover, the presence of enzymes and microorganisms with possible biotechnological applications was confirmed, including proteases, lipases and cell-wall degrading enzymes, pointing to the potential for the hot spring as a source for thermozymes.

Influence of riparian thinning on trophic pathways supporting stream food webs in forested watersheds

AbstractResource managers seek to thin second‐growth riparian forests to address multiple stream and riparian management objectives, including enhancing aquatic productivity via light‐mediated trophic pathways in watersheds of the Pacific Northwest (USA). However, such increases in aquatic productivity depend on complex food web dynamics that link riparian forests and streams. To evaluate how riparian forest thinning influences stream food webs, we conducted a replicated, manipulative field experiment in three northern California watersheds composed of second‐growth redwood forests and tracked responses across multiple trophic levels (periphyton, macroinvertebrates, amphibians, and fish) 1 year pre‐ and post‐treatment. Riparian thinning treatments increased light to the stream channel, yet we observed mixed responses by stream food webs. Thinning did not change stream periphyton biomass on natural substrates but increased periphyton accrual on ceramic tiles. Periphyton accrual appeared to be partially muted by top‐down effects from invertebrate scrapers, which were more abundant in thinned reaches. Prey in the diets of top predators—coastal giant salamanders (Dicamptodon tenebrosus) and coastal cutthroat trout (Oncorhynchus clarkii clarkii)—did not change in biomass, composition, or structure in response to thinning and instead varied more seasonally and between predators. Stable isotope analysis indicated that shifts in carbon (δ13C) signatures of stream periphyton associated with thinning were reflected to varying extents by primary consumers but did not propagate up to top predators. Top predator biomass responses varied between species, where salamander biomass remained unchanged, but cutthroat trout biomass increased slightly in thinned reaches. However, trout biomass responses were not supported by diets or isotopes and correlated weakly with changes in light associated with thinning, suggesting little evidence that responses could be attributed directly to changes in autotrophic pathways. Furthermore, we found no evidence that local trophic responses to thinning propagated into downstream reaches. Taken together, we observed that trophic pathways supporting stream food webs remained largely intact immediately after riparian thinning treatments. Collectively, these results suggest that riparian thinning does not necessarily enhance aquatic productivity in forested streams, indicating that contextual factors driving realized ecological responses should be accounted for when considering thinning as a restoration strategy for stream–riparian ecosystems.

Genomic Features of the Bundle-Forming Heliobacterium Heliophilum fasciatum.

Eight species of heliobacteria have had their genomes sequenced. However, only two of these genomes have been analyzed in detail, those from the thermophilic Heliomicrobium (Hmi.) modesticaldum and the alkaliphilic Heliorestis (Hrs.) convoluta. Here we present analyses of the draft genome sequence of a species of heliobacterium that grows optimally at a moderate temperature and neutral pH. The organism, Heliophilum (Hph.) fasciatum, is phylogenetically unique among cultured heliobacteria and was isolated from rice soil, a common habitat for heliobacteria. The Hph. fasciatum genome contains 3.14 Mbp—similar to that of other reported heliobacteria—but has a G+C base ratio that lies between that of Hmi. modesticaldum and Hrs. convoluta. Many of the genomic features of Hmi. modesticaldum and Hrs. convoluta, such as the absence of genes encoding autotrophic pathways, the presence of a superoperonal cluster of photosynthesis-related genes, and genes encoding endospore-specific proteins, are also characteristic of the Hph. fasciatum genome. However, despite the fact that Hph. fasciatum is diazotrophic, classical nif genes encoding the alpha and beta subunits of dinitrogenase (nifDK) present in other heliobacteria could not be identified. Instead, genes encoding several highly divergent NifDK homologs were present, at least one of which likely encodes a functional dinitrogenase and another a methylthio-alkane reductase (MarDK) for sulfur assimilation. A classical NifH (dinitrogenase reductase) homolog was also absent in Hph. fasciatum, but a related protein was identified that likely carries out this function as well as electron delivery to MarDK. The N2-fixing system of Hph. fasciatum is therefore distinct from that of other heliobacteria and may have unusual properties.

Phylogeny and Metabolic Potential of the Candidate Phylum SAR324.

Simple SummarySAR324, newly proposed as its own candidate phylum, is a diverse and globally abundant bacterial group living in a wide range of environments, from deep-sea hydrothermal vents and brine pools to the epipelagic regions of the global oceans and terrestrial aquifers. The different SAR324 clades harbor a diverse array of genes and pathways well adapted to their respective environments. This metabolic flexibility explains the ubiquitous presence and the importance of SAR324 in global biogeochemical cycles.The bacterial SAR324 cluster is ubiquitous and abundant in the ocean, especially around hydrothermal vents and in the deep sea, where it can account for up to 30% of the whole bacterial community. According to a new taxonomy generated using multiple universal protein-coding genes (instead of the previously used 16S rRNA single gene marker), the former Deltaproteobacteria cluster SAR324 has been classified since 2018 as its own phylum. Yet, very little is known about its phylogeny and metabolic potential. We downloaded all publicly available SAR324 genomes (65) from all natural environments and reconstructed 18 new genomes using publicly available oceanic metagenomic data and unpublished data from the waters underneath the Ross Ice Shelf. We calculated a global SAR324 phylogenetic tree and identified six clusters (namely 1A, 1B, 2A, 2B, 2C and 2D) within this clade. Genome annotation and metatranscriptome read mapping showed that SAR324 clades possess a flexible array of genes suited for survival in various environments. Clades 2A and 2C are mostly present in the surface mesopelagic layers of global oceans, while clade 2D dominates in deeper regions. Our results show that SAR324 has a very versatile and broad metabolic potential, including many heterotrophic, but also autotrophic pathways. While one surface water associated clade (2A) seems to use proteorhodopsin to gain energy from solar radiation, some deep-sea genomes from clade 2D contain the complete Calvin–Benson–Bassham cycle gene repertoire to fix carbon. This, in addition to a variety of other genes and pathways for both oxic (e.g., dimethylsulfoniopropionate degradation) and anoxic (e.g., dissimilatory sulfate reduction, anaerobic benzoate degradation) conditions, can help explain the ubiquitous presence of SAR324 in aquatic habitats.

Autotrophs are important contributors to benthic macroinvertebrate food webs in two tropical first‐order forest streams

Abstract In tropical first‐order streams, the availability of basal resources changes according to tropical seasonality, i.e., dry and rainy seasons, with consequences for the food web structure. However, the seasonal variation in autotrophic and detrital pathways remains unclear. In this study, we aimed to establish whether the dominant pathway of the benthic macroinvertebrate food web in two tropical first‐order streams in the Lacandona rainforest, Mexico, relies on autotrophic or detrital food resources. We evaluated the assimilation of autotrophic and detrital food resources through carbon and nitrogen stable isotope (δ13C and δ15N) analysis. We included the biomass of the macroinvertebrates sustained by the autotrophic and detrital pathways and the food web structure—trophic position, size structure, and degree of omnivory. The biomass sustained by the detrital pathway remained similar throughout the year in both streams. In contrast, the biomass sustained by the autotrophic pathway was higher in the dry season than in the rainy season. The benthic macroinvertebrate food web had four levels, a poorly developed size structure, and an important degree of omnivory in both streams and in both seasons. It is likely that omnivory was a response to high resource variability and a strategy to reduce intraguild competition. Autotrophs had a relatively higher importance than detritus since a much lower algae biomass sustained an equal or higher macroinvertebrate biomass, particularly in the dry season when the biomass of autotrophs increased. Seasonality influenced the macroinvertebrate food web by modifying the food resource availability and transference to higher trophic levels. However, the high frequency of omnivory reduced the seasonal changes in the food web structure. Our results suggest that the autotrophic pathway plays an important role in maintaining the benthic macroinvertebrate food web in tropical streams. The relative importance of autotrophs in the macroinvertebrate food web is associated with seasonality, the rainfall pattern, and its effect on the stream hydrology.

Plants with nitrate preference can regulate nitrification to meet their nitrate demand

Plants with ammonium preference are able to exude biological nitrification inhibitors to reduce nitrification keeping the mineral N in NH4+ form. The question is whether plants with a NO3− preference are able to stimulate nitrification to shift mineral N towards NO3− production to meet their NO3− demand. In this study we attempted to solve this conundrum by conducting 15N tracing studies in a range of soils planted with wheat (Triticum aestivum L.), a typical NO3−-preferring crop, to quantify the gross rates of soil N transformations and the plant N uptake rates. Gross N mineralization rates (M) were stimulated by the presence of wheat in all studied soils, improving the mineral N supply. The wheat NH4+ uptake rates (UNH4) were significantly, positively correlated with M (p < 0.01). The wheat NO3− uptake rates (UNO3) were significantly higher than UNH4 confirming the NO3− preference of this plant. As NO3− production pathways we considered NH4+ oxidation (ONH4, the autotrophic pathway) and organic N oxidation to NO3−, ONrec) in this study. The stimulations of ONH4 were only observed in three out of five soils and, except one soil, ONH4 was much lower (average 1.29 mg N kg−1 d−1) than UNO3 (average 7.66 mg N kg−1 d−1) showing that the NO3− supply via this pathway was insufficient to meet the plants NO3− demand. In these soils, ONrec was significantly stimulated ranging from 0.86 to 5.52 mg N kg−1 d−1 and was responsible for 34%–74% of NO3− production during the 30 days experimental duration. Moreover, UNO3 was significantly, positively correlated with ONrec (p < 0.05), indicating a direct link between heterotrophic nitrification and plant NO3− uptake. One soil (SC2) exhibited a much higher ONH4 (>8.00 mg N kg−1 d−1) and only M was stimulated by the plants presence but not heterotrophic nitrification because the NO3− supply via ONH4 was sufficient to meet the plant NO3− demand. Heterotrophic nitrification was stimulated by NO3− preference plants when NO3− supply via oxidation of NH4+ to NO3− was insufficient to meet the NO3− requirements.

Time-Integrative Multibiomarker Detection in Triassic–Jurassic Rocks from the Atacama Desert: Relevance to the Search for Basic Life Beyond Earth

Detecting evidence of life on other planetary bodies requires a certain understanding of known biomarkers and their chemical nature, preservation potential, or biological specificity. In a planetary search for life, carbonates are of special interest due to their known association with life as we know it. On Earth, carbonates serve as an invaluable paleogeochemical archive of fossils of up to billions of years old. Here, we investigated biomarker profiles on three Chilean Triassic-Jurassic sedimentary records regarding our search for signs of past and present life over ∼200 Ma. A multianalytical platform that combines lipid-derived biomarkers, metaproteomics, and a life detector chip (LDChip) is considered in the detection of biomolecules with different perdurability and source-diagnosis potential. The combined identification of proteins with positive LDChip inmunodetections provides metabolic information and taxonomic affiliation of modern/subrecent biosignatures. Molecular and isotopic analysis of more perdurable hydrocarbon cores allows for the identification of general biosources and dominant autotrophic pathways over time, as well as recreation of prevailing redox conditions over ∼200 Ma. We demonstrate how extraterrestrial life detection can benefit from the use of different biomarkers to overcome diagnosis limitations due to a lack of specificity and/or alteration over time. Our findings have implications for future astrobiological missions to Mars.

Scaling-Up and Long-Term Operation of a Full-Scale Two-Stage Partial Nitritation-Anammox System Treating Landfill Leachate

(1) Background: Biological treatment of leachate in landfill sites using anaerobic ammonium oxidation (anammox) is challenging because of the intrinsic characteristics of this complex wastewater. In this work, the scale-up and subsequent full-scale implementation of the PANAMMOX® technology (LEQUIA Research Group, Girona, Catalonia, Spain) are presented as a case study to achieve long-term nitrogen (N) removal from mature leachate mostly through a completely autotrophic pathway. (2) Methods: The treatment system consists of two sequencing batch reactors (SBRs) running in series to individually operate partial nitritation (PN) and anammox (A). Following biological treatment, physicochemical oxidation (i.e., Fenton-based process) was used to remove the remaining non-biodegradable organic matter. A cost analysis comparative was conducted in relation to the former technology used on-site for treating the leachate. (3) Results: The scale-up of the process from pilot- to full-scale was successfully achieved, finally reaching an average removal of 7.4 kg N/d. The composition of the leachate changed over time, but especially once the landfill site stopped receiving solid waste (this fact involved a marked increase in the strength of the leachate). The adjustment of the alkalinity-to-ammonium ratio before feeding PN-SBR helped to improve the N-removal efficiency. Values of conductivity above 25 mS/cm in A-SBR could negatively affect the performance of the anammox process, making it necessary to consider a dilution strategy according to the on-line monitoring of this parameter. The analysis of the operational costs showed that by implementing the PANAMMOX® technology (LEQUIA Research Group, Girona, Catalonia, Spain) in the landfill site, savings up to 32% were achievable. (4) Conclusions: Treatment of mature landfill leachate in such a two-stage PN-A system was demonstrated as feasible and economically appealing despite the complexity of this industrial wastewater. Accurate expert supervision of the process was a key factor to reaching good performances.

Review: Formation and Metabolic Function of Coral Rubble Biofilms in the Reef Ecosystem

When coral dies, their calcareous skeletons constitute coral rubble in conjunction with the cementing activity of coralline algae and bacteria, creating a secondary reef structure which takes from years to decades to form. Healthy coral reefs differ from coral—rubble dominated reefs in microbial taxonomic composition and metabolic functional roles. The metabolisms of healthy reefs are dominated by autotrophic pathways, where carbon and nitrogen fixation dominate, while the metabolism of rubble—dominated reefs predominate in degradation of organic matter. Nitrogen fixation is 3 orders of magnitude lower in rubble—dominated reefs than in healthy reefs. Coral—rubble harbors a vast diversity of microbes that can precipitate carbonate through coupling several metabolic processes including photosynthesis, ureolysis, ammonification, denitrification, sulfate reduction, methane oxidation, and anaerobic sulfide oxidation. All these metabolic processes were found in rubble microbial communities, but ammonification and sulfate reduction were most prevalent. Anthropogenic and non—anthropogenic perturbations of healthy coral reefs in the past decades have led to the prevalence of rubble—dominated reefs in areas of the Caribbean where the ecological and functional shifts of the community still need further study.

Metagenomic and 14 C tracing evidence for autotrophic microbial CO2 fixation in paddy soils.

Autotrophic carbon dioxide (CO2 ) fixation by microbes is ubiquitous in the environment and potentially contributes to the soil organic carbon (SOC) pool. However, the multiple autotrophic pathways of microbial carbon assimilation and fixation in paddy soils remain poorly characterized. In this study, we combine metagenomic analysis with 14 C-labelling to investigate all known autotrophic pathways and CO2 assimilation mechanisms in five typical paddy soils from southern China. Marker genes of six autotrophic pathways are detected in all soil samples, which are dominated by the cbbL genes (67%-82%) coding the ribulose-bisphosphate carboxylase large chain in the Calvin cycle. These marker genes are associated with a broad range of phototrophic and chemotrophic genera. Significant amounts of 14 C-CO2 are assimilated into SOC (74.3-175.8 mg 14 C kg-1 ) and microbial biomass (5.2-24.1 mg 14 C kg-1 ) after 45 days incubation, where more than 70% of 14 C-SOC was concentrated in the relatively stable humin fractions. These results show that paddy soil microbes contain the genetic potential for autotrophic carbon fixation spreading over broad taxonomic ranges, and can incorporate atmospheric carbon into organic components, which ultimately contribute to the stable SOC pool.

CO2-dependent carbon isotope fractionation in Archaea, Part I: Modeling the 3HP/4HB pathway

The 3-hydroxypropionate/4-hydroxybutyrate (3HP/4HB) pathway of carbon fixation is found in thermophilic Crenarchaeota of the order Sulfolobales and in aerobic, ammonia-oxidizing Thaumarchaeota. Unlike all other known autotrophic carbon metabolisms, this pathway exclusively uses HCO_3- rather than CO_2 as the substrate for carbon fixation. Biomass produced by the 3HB/4HP pathway is relatively ^(13)C-enriched compared to biomass fixed by other autotrophic pathways, with total biosynthetic isotope effects (e_(Ar)) of ca. 3‰ in the Sulfolobales and ca. 20‰ in the Thaumarchaeota. Explanations for the difference between these values usually invoke the dual effects of thermophily and growth at low pH (low [HCO_3-]) for the former group vs. mesophily and growth at pH > 7 (high [HCO_3-]) for the latter group. Here we examine the model taxa Metallosphaera sedula and Nitrosopumilus maritimususing an isotope flux-balance model to argue that the primary cause of different e_(Ar) values more likely is the presence of carbonic anhydrase in M. sedula and its corresponding absence in N. maritimus. The results suggest that the pool of HCO_3-inside N. maritimus is out of isotopic equilibrium with CO_2 and that the organism imports < 10% HCO_3- from the extracellular environment. If correct and generalizable, the aerobic, ammonia-oxidizing marine Thaumarchaeota are dependent on passive CO_2 uptake and a slow rate of intracellular conversion to HCO_3-. Values of e_(Ar) should therefore vary in response to growth rate (μ) and CO_2 availability, analogous to eukaryotic algae, but in the opposite direction: e_(Ar) becomes smaller as [CO_(2(aq))] increases and/or μ decreases. Such an idea represents a testable hypothesis, both in the laboratory and in natural systems. Sensitivity to μ and CO_2 implies that measurements of e_(Ar) may hold promise as a pCO_2 paleobarometer.

Sulfur-Oxidizing Symbionts without Canonical Genes for Autotrophic CO2 Fixation.

Since the discovery of symbioses between sulfur-oxidizing (thiotrophic) bacteria and invertebrates at hydrothermal vents over 40 years ago, it has been assumed that autotrophic fixation of CO2 by the symbionts drives these nutritional associations. In this study, we investigated "Candidatus Kentron," the clade of symbionts hosted by Kentrophoros, a diverse genus of ciliates which are found in marine coastal sediments around the world. Despite being the main food source for their hosts, Kentron bacteria lack the key canonical genes for any of the known pathways for autotrophic carbon fixation and have a carbon stable isotope fingerprint that is unlike other thiotrophic symbionts from similar habitats. Our genomic and transcriptomic analyses instead found metabolic features consistent with growth on organic carbon, especially organic and amino acids, for which they have abundant uptake transporters. All known thiotrophic symbionts have converged on using reduced sulfur to gain energy lithotrophically, but they are diverse in their carbon sources. Some clades are obligate autotrophs, while many are mixotrophs that can supplement autotrophic carbon fixation with heterotrophic capabilities similar to those in Kentron. Here we show that Kentron bacteria are the only thiotrophic symbionts that appear to be entirely heterotrophic, unlike all other thiotrophic symbionts studied to date, which possess either the Calvin-Benson-Bassham or the reverse tricarboxylic acid cycle for autotrophy.IMPORTANCE Many animals and protists depend on symbiotic sulfur-oxidizing bacteria as their main food source. These bacteria use energy from oxidizing inorganic sulfur compounds to make biomass autotrophically from CO2, serving as primary producers for their hosts. Here we describe a clade of nonautotrophic sulfur-oxidizing symbionts, "Candidatus Kentron," associated with marine ciliates. They lack genes for known autotrophic pathways and have a carbon stable isotope fingerprint heavier than other symbionts from similar habitats. Instead, they have the potential to oxidize sulfur to fuel the uptake of organic compounds for heterotrophic growth, a metabolic mode called chemolithoheterotrophy that is not found in other symbioses. Although several symbionts have heterotrophic features to supplement primary production, in Kentron they appear to supplant it entirely.

Methanogenesis on Early Stages of Life: Ancient but Not Primordial.

Of the six known autotrophic pathways, the Wood-Ljungdahl pathway (WL) is the only one present in both the acetate producing Bacteria (homoacetogens) and the methane producing Archaea (hydrogenotrophic methanogens), and it has been suggested that WL is one of the oldest metabolic pathways. However, only the so-called carbonyl branch is shared by Archaea and Bacteria, while the methyl branch is different, both in the number of reactions and enzymes, which are not homologous among them. In this work we show that some parts of the methyl branch of archaeal Wood-Ljungdahl pathway (MBWL) are present in bacteria as well as in non-methanogen archaea, although the tangled evolutionary history of MBWL cannot be traced back to the Last Common Ancestor. We have also analyzed the different variants of methanogenesis (hydrogenotrophic, acetoclastic and methylotrophic pathways), and concluded that each of these pathways, and every different enzyme or subunit (in the case of multimeric enzymes), has their own intricate evolutionary history. Our study supports the scenario of hydrogenotrophic methanogenesis being older than the other variants, albeit not old enough to be present in the last archaeal common ancestor.