Matching Items (25)

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Complete Genome Sequence of Methanosphaerula palustris E1-9CT, a Hydrogenotrophic Methanogen Isolated from a Minerotrophic Fen Peatland

Description

Here, we report the complete genome sequence (2.92 Mb) of Methanosphaerula palustris E1-9C[superscript T], a methanogen isolated from a minerotrophic fen. This is the first genome report of the Methanosphaerula

Here, we report the complete genome sequence (2.92 Mb) of Methanosphaerula palustris E1-9C[superscript T], a methanogen isolated from a minerotrophic fen. This is the first genome report of the Methanosphaerula genus, within the Methanoregulaceae family, in the Methanomicrobiales order. E1-9C[superscript T] relatives are found in a wide range of ecological and geographical settings.

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  • 2015-11-05

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Exometabolite niche partitioning among sympatric soil bacteria

Description

Soils are arguably the most microbially diverse ecosystems. Physicochemical properties have been associated with the maintenance of this diversity. Yet, the role of microbial substrate specialization is largely unexplored since

Soils are arguably the most microbially diverse ecosystems. Physicochemical properties have been associated with the maintenance of this diversity. Yet, the role of microbial substrate specialization is largely unexplored since substrate utilization studies have focused on simple substrates, not the complex mixtures representative of the soil environment. Here we examine the exometabolite composition of desert biological soil crusts (biocrusts) and the substrate preferences of seven biocrust isolates. The biocrust's main primary producer releases a diverse array of metabolites, and isolates of physically associated taxa use unique subsets of the complex metabolite pool. Individual isolates use only 13−26% of available metabolites, with only 2 out of 470 used by all and 40% not used by any. An extension of this approach to a mesophilic soil environment also reveals high levels of microbial substrate specialization. These results suggest that exometabolite niche partitioning may be an important factor in the maintenance of microbial diversity.

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  • 2015-09-22

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Photosynthesis under Rocks: Hypolith Distribution across the Namib Desert Rainfall Gradient

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“Extremophile” is used to describe life that has adapted to extreme conditions and the conditions they live in are often used to understand the limits of life. In locations with

“Extremophile” is used to describe life that has adapted to extreme conditions and the conditions they live in are often used to understand the limits of life. In locations with low precipitation and high solar radiation, photosynthetic cyanobacteria can colonize the underside of quartz fragments, forming ‘hypoliths.’ The quartz provides protection against wind, reduces solar radiation, and slows the rate of evaporation following infrequent rain or fog events. In most desert systems, vascular plants are the main primary producers. However, hypoliths might play a key role in carbon fixation in hyperarid deserts that are mostly devoid of vegetation. I investigated hypolith distribution and carbon fixation at six sites along a rainfall and fog gradient in the central Namib Desert in Namibia. I used line point intersect transects to assess ground cover (bare soil, colonized quartz fragment, non-colonized quartz fragment, non-quartz rock, grass, or lichen) at each site. Additionally, at each site I selected 12 hypoliths and measured cyanobacteria colonization on quartz and measured CO2 flux of hypoliths at five of the six sites.
Ground cover was fairly similar among sites, with bare ground > non-colonized quartz fragments > colonized quartz fragments > non-quartz rocks. Grass was present only at the site with the highest mean annual precipitation (MAP) where it accounted for 1% of ground cover. Lichens were present only at the lowest MAP site, where they accounted for 30% of ground cover. The proportion of quartz fragments colonized generally increased with MAP, from 5.9% of soil covered by colonized hypoliths at the most costal (lowest MAP) site, to 18.7% at the most inland (highest MAP) site. There was CO2 uptake from most hypoliths measured, with net carbon uptake rates ranging from 0.3 to 6.4 μmol m-2 s-1 on well hydrated hypoliths. These carbon flux values are similar to previous work in the Mojave Desert. Our results suggest that hypoliths might play a key role in the fixation of organic carbon in hyperarid ecosystems where quartz fragments are abundant, with MAP constraining hypolith abundance. A better understanding of these extremophiles and the niche they fill could give an understanding of how microbial life might exist in extraterrestrial environments similar to hyperarid deserts.

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  • 2016-12

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Trait-Based Modeling of Peatland Methanogen Communities

Description

Methanogens are methane-producing archaea that play a major role in the global carbon cycle. However, despite their importance, the community dynamics of these organisms have not been thoroughly characterized or

Methanogens are methane-producing archaea that play a major role in the global carbon cycle. However, despite their importance, the community dynamics of these organisms have not been thoroughly characterized or modeled. In the majority of methanogenesis models, the communities are approximated as a chemical reaction or divided into two populations based on the most common methanogenic pathways. These models provide reasonable estimate of methanogenesis rates but cannot predict community structure. In this work, a trait-based model for methanogenic communities in peatlands is developed. The model divides methanogens commonly found in wetlands into ten guilds, with divisions based on factors such as substrate affinity, pH tolerance, and phylogeny. The model uses steady-state, mixotrophic Monod kinetics to model growth and assumes peatlands operate as a semi-batch system. An extensive literature review was performed to parameterize the model. The acetoclastic module of the model was validated against experimental data. It was found that this portion of the model was able to reproduce the major result of an experiment that examined competition between Methanosaeta and Methanosarcina species under irregular feeding conditions. The model was analyzed as a whole using Monte Carlo simulation methods. It was found that equilibrium membership is negatively correlated with a guild's half-substrate constant, but independent of the guild's yield. These results match what is seen in simple pairwise competition models. In contrast, it was found that both the half-substrate constant and yield affected a guild's numerical dominance. Lower half-substrate constants and higher yields led to a guild accounting for a greater fraction of community biomass. This is not seen in simple pairwise competitions models where only yield affects final biomass. As a whole, the development of this model framework and the accompanying analyses have laid the groundwork for a new class of more detailed methanogen community models that go beyond the two compartment acetoclastic-hydrogenotrophic assumption. .

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  • 2017-05

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Over-expression of a putative multi-heme cytochrome c from Heliobacterium modesticaldum

Description

Heliobacterium modesticaldum (H. modesticaldum) is an anaerobic photoheterotroph that can fix nitrogen (N2) and produce molecular hydrogen (H2). Recently, the Redding and Jones labs created a microbial photoelectrosynthesis cell that

Heliobacterium modesticaldum (H. modesticaldum) is an anaerobic photoheterotroph that can fix nitrogen (N2) and produce molecular hydrogen (H2). Recently, the Redding and Jones labs created a microbial photoelectrosynthesis cell that utilized these properties to produce molecular hydrogen using electrons provided by a cathode via a chemical mediator. Although this light-driven creation of fuel within a microbial electrochemical cell was the first of its kind, its production rate of hydrogen was low. It was hypothesized that the injection of electrons into H. modesticaldum was a rate-limiting step in H2 production. Within the H. modesticaldum genome, there is a gene (HM1_0653) that encodes a multi-heme cytochrome c that may be directly involved in this step. From past transcriptomic experiments, this gene is known to be very poorly expressed in H. modesticaldum. Our hypothesis was that increasing its expression with a strong promoter could result in faster electron transfer, and thus, increased H2 production in the photoelectrosynthesis cell. In order to test this hypothesis, different promoters that could lead to high expression in H. modesticaldum were included with a copy of HM1_0653 in various plasmid constructs that were first cloned into E. coli before being conjugated with H. modesticaldum. Cloning in E. coli was possible with the newly derived transformation system and by reducing the copy-number of the vector system. When overexpressed in E. coli, the protein appeared to be expressed, but its purification proved to be difficult. Moreover, conjugation with H. modesticaldum was not achieved. Our results are consistent with the idea that high level overexpression in H. modesticaldum was toxic. An inducible promoter may circumvent these issues and prove more successful in future experiments.

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Date Created
  • 2018-05

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Assessment of the Vertical Stratification of Microbial Community Structure in Permafrost Peatlands

Description

Peatlands are a type of wetlands where the rate of accumulation of organic matter exceed the rate of decomposition and have accumulated more than 30 cm of peat (Joosten and

Peatlands are a type of wetlands where the rate of accumulation of organic matter exceed the rate of decomposition and have accumulated more than 30 cm of peat (Joosten and Clark, 2002). Peatlands store approximately 30% of all terrestrial carbon as recalcitrant peat, partially decomposed plant and microbial biomass, while simultaneously producing almost 40% of the globally emitted methane (Schmidt et al., 2016), making peatlands an important component of the carbon budgets. Published research indicates that the efficiency of carbon usage among microbial communities can determine the soil-carbon response to rising temperatures (Allison et al. 2010). By determining carbon consumption in peatland soils, total community respiration response, and community structure change with additions, models of carbon use efficiency in permafrost peatlands will be well-informed and have a better understanding of how the peatlands will respond to, and utilize, increased availability of carbon compounds due to the melting permafrost. To do this, we will sequence Lutose deep core samples to observe baseline microbial community structure at different depths and different age-gradients, construct substrate incubations of glucose and propionate and observe community respiration response via a gas chromatography flame ionization detector, track the glucose and propionate additions with high-performance liquid chromatography (HPLC), and sequence the samples once more to determine if there was a deviation from the initial community structure obtained prior to the incubations. We found that our initial sequencing data was supported by previous work (Lin et al., 2014), however we were unable to sequence samples post-incubation due to time constraints. In this sequencing analysis we found that the strongest variable that made samples biologically similar was the age-gradient site in which they were extracted. We found that the group with glucose additions produced the most carbon dioxide compared with the other treatments, but was not the treatment that dominated the production of methane. Finally, in the HPLC samples that were analyzed, we found that glucose is likely forming the most by-product accumulation from mass balance calculations, while propionate is likely forming the least. Future experimentation should focus on the shortcomings of this experiment. Further analysis of 16S rRNA sequencing data from after the incubations should be analyzed to determine the change in microbial community structure throughout the experiment. Furthermore, HPLC analysis for the several samples need to be done and followed up with mass balance to determine where the added glucose and propionate are being allocated within the soil. Once these pieces of the puzzle are put into place, our original question of how the microbial community structure changes at different depths and age-gradients within permafrost peatlands will be conclusively answered.

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  • 2018-05

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Marine Aggregation interactions of Prochlorococcus and Marinobacter adhaerens

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The changes in marine ecological conditions brought on by warming and stratification of the oceans have radically shifted many marine environments around the globe. This project aimed to better characterize

The changes in marine ecological conditions brought on by warming and stratification of the oceans have radically shifted many marine environments around the globe. This project aimed to better characterize the aggregation behavior of the abundant picocyanobacterium Prochlorococcus marinus, which is hypothesized to dominate over other phytoplankton as the primary autotroph in increasingly warmer and nutrient poor oceans. This aggregation, believed to be mediated through the secretion of sticky Transparent Exopolymeric Substances (TEP), might be key for Prochlorococcus to sink throughout the ocean and serve as a source of carbon to other communities within its environment. Considering the relatively low concentration of TEP secreted by Prochlorococcus when on its own, this study explored the synergistic effect that heterotrophic bacteria and inorganic minerals in the surrounding seawater may have on the aggregation of P. marinus. This was done by inoculating P. marinus and the model heterotroph Marinobacter adhaerens HP15 individually and mixed in cylindrical roller tanks with the addition of ballasting clay minerals into roller tanks to simulate constant sinking for 7 days. The aggregates which formed after rolling were quantified and their sinking velocities and excess densities were measured. Our results indicate that the most numerous and densest aggregates formed when Prochlorococcus was in the presence of both M. adhaerens and kaolinite clay particles. I will discuss how methodology, particularly cell number, may play a role in the enhanced aggregation that I found when Prochlorococcus was cultured together with the Marinobacter.

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  • 2020-05

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Evaluating the Consumption Rates of Primary Versus Secondary Fermentation Substrates and Methane Production of Northern Peatlands

Description

Northern peatland carbon cycling is under close observation and is critical to include in models projecting the future effects of climate change as these ecosystems represent a significant source of

Northern peatland carbon cycling is under close observation and is critical to include in models projecting the future effects of climate change as these ecosystems represent a significant source of atmospheric methane (CH4). Changes in the in situ conditions, brought upon by the warming climate, could alter the rates of organic matter decomposition and accelerate the emissions of greenhouse, changing northern peatland’s status as a carbon sink. In order to develop a better understanding of the climate’s effect on the microbial community composition, carbon decomposition cascade, and flux of CH4 and CO2, anoxic soil microcosms were supplemented with either glucose or propionate to test the distinct intermediary metabolism of four northern peatland sites with statistically similar geochemistry that exist across a climate gradient. Lutose (LT) and Bog Lake (BL) consumed the supplemented glucose at the highest rates, 42.6 mg/L per day and 39.5 mg/L per day respectively. Chicago Bog (CB) and Daring Lake (DL) consumed the supplemented propionate at the highest rates, 5.26 mg/L per day and 4.34 mg/L per day respectively. BL microcosms showed low levels of methanogenesis as CH4 concentrations reached a maximum of 2.61 µmol/g dry soil in the treatments. In DL, the site with the highest production of CH4, the low abundance of hydrogenotrophic methanogens (Methanocellaceae and Methanoregulaceae) and relatively steady concentrations of acetate and formate could indicate that these are the more desired methanogenic substrates. These findings are indicative of the differences in metabolic potential found across these geochemically similar peatlands, lending to climate variables being a major driver in microbial community potential. To further characterize the intermediary metabolism and the effect of the climate gradient in these sites, future experimentations should incorporate 13C DNA-stable isotope probing data, establish a mass balance of the system, and incubate the microcosms at their respective in situ temperatures.

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  • 2020-05

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Qualitative Determination of Exopolymeric Substances in Particle-Associated Bacteria from the Sargasso Sea

Description

The biological carbon pump acts as part of the global carbon cycle through the photosynthetic fixation of inorganic carbon into dissolved and particulate organic carbon by phytoplankton. Previously, the biological

The biological carbon pump acts as part of the global carbon cycle through the photosynthetic fixation of inorganic carbon into dissolved and particulate organic carbon by phytoplankton. Previously, the biological carbon pump was attributed to large aggregates and zooplankton fecal pellets since their size and density results in faster sinking rates, efficiently exporting organic carbon to deeper depths in the ocean. However, recent studies have indicated that small cells, known as picoplankton, contribute significantly to the formation of sinking particles. The presence of exopolymeric substances (EPS), among them sticky transparent exopolymeric particles (TEP) and proteinaceous coomassie stainable particles (CSP), serve as influential factors of export flux and aggregation. The presence of heterotrophic bacteria can also affect aggregation and sinking velocity, as seen in previous studies, and is likely attributed to their EPS and TEP production. The staining and visualization of TEP and CSP allow for the qualitative determination of these types of EPS from bacteria isolated from sinking particles collected with particle interceptor traps at various depths in the Sargasso Sea. I study the presence of TEP and CSP in particle-associated bacteria. Cultures of picocyanobacteria, consisting of xenic Synechococcus and axenic Prochlorococcus, were used to establish positive and negative controls for stained isolate analysis. Marinobacter adhaerens served as a tertiary control for an axenic culture that stains positive for TEP. I chose six isolates of bacteria isolated from sinking particles to be stained and visualized to test for the secretion of TEP and CSP. Four of the isolates stained positive for both TEP and CSP, including Pseudoalteromonas sp., Erythrobacter sp., and Marinobacter sp., while one isolate, Micrococcus sp., stained positive only for TEP, and the last isolate, another Marinobacter sp., stained positive for only CSP. These results are important in understanding the role of plankton organisms in the formation of sinking particles.

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Date Created
  • 2021-05

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Evaluating Drivers of Chemodenitrification in Tropical Peat Soil

Description

Nitrous oxide (N2O) is a major contributor to the greenhouse effect and to stratospheric ozone depletion. In soils, nitrogen reduction is performed by biotic and abiotic processes, including microbial denitrification

Nitrous oxide (N2O) is a major contributor to the greenhouse effect and to stratospheric ozone depletion. In soils, nitrogen reduction is performed by biotic and abiotic processes, including microbial denitrification and chemical denitrification. Chemical denitrification, or chemodenitrification, is the abiotic step-wise reduction of nitrate (NO3-), nitrite (NO2-), or nitric oxide (NO) to N2O in anoxic environments, with high turnover rates particularly in acidic soils. Chemodenitrification was identified in various environments, but the mechanism is still not understood. In this study, the factors influencing abiotic reduction of NO2- to N2O in acidic tropical peat soil are examined. These factors include pH, organic matter content, and dissolved ferrous iron. Anoxic peat soil from sites located in the Peruvian Amazon was used for incubations. The results show that peat soil (pH ~4.5) appears to reduce NO2- more quickly in the presence of lower pH and higher Fe(II) concentrations. NO2- is completely reduced in excess Fe(II), and Fe(II) is completely oxidized in excess NO2-, providing evidence for the proposed mechanism of chemodenitrification. In addition, first order reaction rate constants kFe(II) and kNO2- were calculated using concentration measurements over 4 hours, to test for the hypothesized reaction rate relationships kFe(II): kFe(II) kFe(II)~NO2- > kFe(II)>NO2- and kNO2-: kFe(II)NO2-. The NO2- k values followed the anticipated pattern, although the Fe(II) k value data was inconclusive. Organic material may also play a role in NO2- reduction through chemodenitrification, and future experimentation will test this possibility. How and to what extent the pH and the concentrations of organic matter and Fe(II) affect the kinetic rate of chemodenitrification will lend insight into the N2O production potential of natural tropical peatlands.

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Date Created
  • 2016-05