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1.
mSystems ; 9(1): e0096623, 2024 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-38059636

RESUMO

Microbes can be found in abundance many kilometers underground. While microbial metabolic capabilities have been examined across different geochemical settings, it remains unclear how changes in subsurface niches affect microbial needs to sense and respond to their environment. To address this question, we examined how microbial extracellular sensor systems vary with environmental conditions across metagenomes at different Deep Mine Microbial Observatory (DeMMO) subsurface sites. Because two-component systems (TCSs) directly sense extracellular conditions and convert this information into intracellular biochemical responses, we expected that this sensor family would vary across isolated oligotrophic subterranean environments that differ in abiotic and biotic conditions. TCSs were found at all six subsurface sites, the service water control, and the surface site, with an average of 0.88 sensor histidine kinases (HKs) per 100 genes across all sites. Abundance was greater in subsurface fracture fluids compared with surface-derived fluids, and candidate phyla radiation (CPR) bacteria presented the lowest HK frequencies. Measures of microbial diversity, such as the Shannon diversity index, revealed that HK abundance is inversely correlated with microbial diversity (r2 = 0.81). Among the geochemical parameters measured, HK frequency correlated most strongly with variance in dissolved organic carbon (r2 = 0.82). Taken together, these results implicate the abiotic and biotic properties of an ecological niche as drivers of sensor needs, and they suggest that microbes in environments with large fluctuations in organic nutrients (e.g., lacustrine, terrestrial, and coastal ecosystems) may require greater TCS diversity than ecosystems with low nutrients (e.g., open ocean).IMPORTANCEThe ability to detect extracellular environmental conditions is a fundamental property of all life forms. Because microbial two-component sensor systems convert information about extracellular conditions into biochemical information that controls their behaviors, we evaluated how two-component sensor systems evolved within the deep Earth across multiple sites where abiotic and biotic properties vary. We show that these sensor systems remain abundant in microbial consortia at all subterranean sampling sites and observe correlations between sensor system abundances and abiotic (dissolved organic carbon variation) and biotic (consortia diversity) properties. These results suggest that multiple environmental properties may drive sensor protein evolution and highlight the need for further studies of metagenomic and geochemical data in parallel to understand the drivers of microbial sensor evolution.


Assuntos
Matéria Orgânica Dissolvida , Ecossistema , Bactérias/genética , Metagenoma , Meio Ambiente
2.
ACS Synth Biol ; 12(12): 3743-3753, 2023 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-37991716

RESUMO

Soil microbial communities with reduced complexity are emerging as model systems for studying consortia-scale phenotypes. To establish synthetic biology tools for studying these communities in hard-to-image environmental materials, we evaluated whether a single member of a model soil consortium (MSC) can be programmed to report on gene expression without requiring matrix disruption. For these studies, we targeted a five-membered MSC that includes Dyadobacter fermentans, Ensifer adhaerens, Rhodococcus sp003130705, Streptomyces sp001905665, and Variovorax beijingensis. By coupling the expression of a methyl halide transferase to a constitutive promoter, we show that V. beijingensis can be programmed to synthesize methyl halides that accumulate in the soil headspace at levels that are ≥24-fold higher than all other MSC members across a range of environmentally relevant hydration conditions. We find that methyl halide production can report on an MSC promoter that is activated by changes in water potential, and we demonstrate that a synthetic gas signal can be read out directly using gas chromatography and indirectly using a soil-derived Methylorubrum that is programmed to produce a visual output in response to methyl halides. These tools will be useful for future studies that investigate how MSC responds to dynamic hydration conditions, such as drought and flood events induced by climate change, which can alter soil water potential and induce the release of stored carbon.


Assuntos
Hidrocarbonetos Bromados , Solo , Solo/química , Água , Transdução de Sinais
3.
Environ Sci Technol ; 57(1): 128-138, 2023 01 10.
Artigo em Inglês | MEDLINE | ID: mdl-36525597

RESUMO

In situ aging can change biochar properties, influencing their ecosystem benefits or risks over time. However, there is a lack of field verification of laboratory methods that attempt simulation of long-term natural aging of biochar. We exploited a decade-scale natural charcoal (a proxy for biochar) aging event to determine which lab-aging methods best mimicked field aging. We oxidized charcoal by ultraviolet A radiation (UVA), H2O2, or monochloramine (NH2Cl), and compared it to 10-year field-aged charcoal. We considered seven selected charcoal properties related to surface chemistry and organic matter release, and found that oxidation with 30% H2O2 most representatively simulated 10-year field aging for six out of seven properties. UVA aging failed to approximate oxidation levels while showing a distinctive dissolved organic carbon (DOC) release pattern. NH2Cl-aged charcoal was the most different, showing an increased persistent free radical (PFR) concentration and lower hydrophilicity. All lab oxidation techniques overpredicted polycyclic aromatic hydrocarbon release. The O/C ratio was well-correlated with DOC release, PFR concentration, surface charge, and charcoal pH, indicating the possibility to accurately predict biochar aging with a reduced suite of physicochemical properties. Overall, our rapid and verified lab-aging methods facilitate research toward derisking and enhancing long-term benefits of biochar application.


Assuntos
Carvão Vegetal , Poluentes do Solo , Carvão Vegetal/química , Solo/química , Ecossistema , Peróxido de Hidrogênio
4.
ACS Synth Biol ; 11(9): 2909-2916, 2022 09 16.
Artigo em Inglês | MEDLINE | ID: mdl-35961652

RESUMO

Microbial biosensors sense and report exposures to stimuli, thereby facilitating our understanding of environmental processes. Successful design and deployment of biosensors hinge on the persistence of the microbial host of the genetic circuit, termed the chassis. However, model chassis organisms may persist poorly in environmental conditions. In contrast, non-model organisms persist better in environmental conditions but are limited by other challenges, such as genetic intractability and part unavailability. Here we identify ecological, metabolic, and genetic constraints for chassis development and propose a conceptual framework for the systematic selection of environmental biosensor chassis. We identify key challenges with using current model chassis and delineate major points of conflict in choosing the most suitable organisms as chassis for environmental biosensing. This framework provides a way forward in the selection of biosensor chassis for environmental synthetic biology.


Assuntos
Técnicas Biossensoriais , Biologia Sintética , Engenharia Metabólica
5.
mSystems ; 7(4): e0030122, 2022 08 30.
Artigo em Inglês | MEDLINE | ID: mdl-35880897

RESUMO

Soil matrix properties influence microbial behaviors that underlie nutrient cycling, greenhouse gas production, and soil formation. However, the dynamic and heterogeneous nature of soils makes it challenging to untangle the effects of different matrix properties on microbial behaviors. To address this challenge, we developed a tunable artificial soil recipe and used these materials to study the abiotic mechanisms driving soil microbial growth and communication. When we used standardized matrices with varying textures to culture gas-reporting biosensors, we found that a Gram-negative bacterium (Escherichia coli) grew best in synthetic silt soils, remaining active over a wide range of soil matric potentials, while a Gram-positive bacterium (Bacillus subtilis) preferred sandy soils, sporulating at low water potentials. Soil texture, mineralogy, and alkalinity all attenuated the bioavailability of an acyl-homoserine lactone (AHL) signaling molecule that controls community-level microbial behaviors. Texture controlled the timing of AHL sensing, while AHL bioavailability was decreased ~105-fold by mineralogy and ~103-fold by alkalinity. Finally, we built artificial soils with a range of complexities that converge on the properties of one Mollisol. As artificial soil complexity increased to more closely resemble the Mollisol, microbial behaviors approached those occurring in the natural soil, with the notable exception of organic matter. IMPORTANCE Understanding environmental controls on soil microbes is difficult because many abiotic parameters vary simultaneously and uncontrollably when different natural soils are compared, preventing mechanistic determination of any individual soil parameter's effect on microbial behaviors. We describe how soil texture, mineralogy, pH, and organic matter content can be varied individually within artificial soils to study their effects on soil microbes. Using microbial biosensors that report by producing a rare indicator gas, we identify soil properties that control microbial growth and attenuate the bioavailability of a diffusible chemical used to control community-level behaviors. We find that artificial soils differentially affect signal bioavailability and the growth of Gram-negative (Escherichia coli) and Gram-positive (Bacillus subtilis) microbes. These artificial soils are useful for studying the mechanisms that underlie soil controls on microbial fitness, signaling, and gene transfer.


Assuntos
Acil-Butirolactonas , Solo , Solo/química , Escherichia coli/genética , Areia , Microbiologia do Solo
6.
ACS Synth Biol ; 11(7): 2372-2383, 2022 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-35715210

RESUMO

Gene expression can be monitored in hard-to-image environmental materials using gas-reporting biosensors, but these outputs have only been applied in autoclaved matrices that are hydrated with rich medium. To better understand the compatibility of indicator gas reporting with environmental samples, we evaluated how matrix hydration affects the gas signal of an engineered microbe added to a sieved soil. A gas-reporting microbe presented a gas signal in a forest soil (Alfisol) when hydrated to an environmentally relevant osmotic pressure. When the gas signal was concentrated prior to analysis, a biosensor titer of 103 cells/gram of soil produced a significant signal when soil was supplemented with halides. A signal was also observed without halide amendment, but a higher cell titer (106 cells/gram of soil) was required. A sugar-regulated gas biosensor was able to report with a similar level of sensitivity when added to an unsterilized soil matrix, illustrating how gas concentration enables biosensing within a soil containing environmental microbes. These results establish conditions where engineered microbes can report on gene expression in living environmental matrices with decreased perturbation of the soil environment compared to previously reported approaches, using biosensor titers that are orders of magnitude lower than the number of cells typically observed in a gram of soil.


Assuntos
Técnicas Biossensoriais , Solo
7.
Sci Rep ; 12(1): 1002, 2022 01 19.
Artigo em Inglês | MEDLINE | ID: mdl-35046439

RESUMO

The costs of COVID-19 are extensive, and, like the fallout of most health and environmental crises in the US, there is growing evidence that these costs weigh disproportionately on communities of color. We investigated whether county-level racial composition and fine particulate pollution (PM2.5) are indicators for COVID-19 incidence and death rates in the state of Texas. Using county-level data, we ran linear regressions of percent minority as well as historic 2000-2016 PM2.5 levels against COVID-19 cases and deaths per capita. We found that a county's percent minority racial composition, defined as the percentage of population that identifies as Black or Hispanic, highly correlates with COVID-19 case and death rates. Using Value-of-Statistical-Life calculations, we found that economic costs from COVID-19 deaths fall more heavily on Black and Hispanic residents in Harris County, the most populous county in Texas. We found no consistent evidence or significant correlations between historic county-average PM2.5 concentration and COVID-19 incidence or death. Our findings suggest that public health and economic aid policy should consider the racially-segregated burden of disease to better mitigate costs and support equity for the duration and aftermath of health crises.


Assuntos
Poluentes Atmosféricos/efeitos adversos , COVID-19/mortalidade , Minorias Étnicas e Raciais/estatística & dados numéricos , Material Particulado/efeitos adversos , SARS-CoV-2/isolamento & purificação , Adolescente , Adulto , Idoso , COVID-19/epidemiologia , COVID-19/virologia , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Prognóstico , Taxa de Sobrevida , Texas/epidemiologia , Adulto Jovem
8.
Eco Environ Health ; 1(4): 212-218, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38077255

RESUMO

The concentration and molecular composition of soil organic matter (SOM) are important factors in mitigation against climate change as well as providing other ecosystem services. Our quantitative understanding of how land use influences SOM molecular composition and associated turnover dynamics is limited, which underscores the need for high-throughput analytical approaches and molecular marker signatures to clarify this etiology. Combining a high-throughput untargeted mass spectrometry screening and molecular markers, we show that forest, farmland and urban land uses result in distinct molecular signatures of SOM in the Lake Chaohu Basin. Molecular markers indicate that forest SOM has abundant carbon contents from vegetation and condensed organic carbon, leading to high soil organic carbon (SOC) concentration. Farmland SOM has moderate carbon contents from vegetation, and limited content of condensed organic carbon, with SOC significantly lower than that of forest soils. Urban SOM has high abundance of condensed organic carbon markers due to anthropogenic activities but relatively low in markers from vegetation. Consistently, urban soils have the highest black carbon/SOC ratio among these land uses. Overall, our results suggested that the molecular signature of SOM varies significantly with land use in the Lake Chaohu Basin, influencing carbon dynamics. Our strategy of molecular fingerprinting and marker discovery is expected to enlighten further research on SOM molecular signatures and cycling dynamics.

9.
ACS Synth Biol ; 9(11): 3104-3113, 2020 11 20.
Artigo em Inglês | MEDLINE | ID: mdl-33104325

RESUMO

Monitoring microbial reactions in highly opaque or autofluorescent environments like soils, seawater, and wastewater remains challenging. To develop a simple approach for observing post-translational reactions within microbes situated in environmental matrices, we designed a methyl halide transferase (MHT) fragment complementation assay that reports by synthesizing an indicator gas. We show that backbone fission within regions of high sequence variability in the Rossmann domain yields split MHT (sMHT) AND gates whose fragments cooperatively associate to synthesize CH3Br. Additionally, we identify a sMHT whose fragments require fusion to pairs of interacting partner proteins for maximal activity. We also show that sMHT fragments fused to FKBP12 and the FKBP-rapamycin binding domain of mTOR display significantly enhanced CH3Br production in the presence of rapamycin. This gas production is reversed in the presence of the competitive inhibitor of FKBP12/FKPB dimerization, indicating that sMHT is a reversible reporter of post-translational reactions. This sMHT represents the first genetic AND gate that reports on protein-protein interactions via an indicator gas. Because indicator gases can be measured in the headspaces of complex environmental samples, this assay should be useful for monitoring the dynamics of diverse molecular interactions within microbes situated in hard-to-image marine and terrestrial matrices.


Assuntos
Gases/metabolismo , Transferases/genética , Dimerização , Pentosiltransferases/genética , Mapas de Interação de Proteínas/efeitos dos fármacos , Mapas de Interação de Proteínas/genética , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Processamento de Proteína Pós-Traducional/genética , Sirolimo/farmacologia , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/farmacologia , Proteína 1A de Ligação a Tacrolimo/genética
10.
Sci Adv ; 6(5): eaax8254, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-32064339

RESUMO

Plant-microbe interactions are mediated by signaling compounds that control vital plant functions, such as nodulation, defense, and allelopathy. While interruption of signaling is typically attributed to biological processes, potential abiotic controls remain less studied. Here, we show that higher organic carbon (OC) contents in soils repress flavonoid signals by up to 70%. Furthermore, the magnitude of repression is differentially dependent on the chemical structure of the signaling molecule, the availability of metal ions, and the source of the plant-derived OC. Up to 63% of the signaling repression occurs between dissolved OC and flavonoids rather than through flavonoid sorption to particulate OC. In plant experiments, OC interrupts the signaling between a legume and a nitrogen-fixing microbial symbiont, resulting in a 75% decrease in nodule formation. Our results suggest that soil OC decreases the lifetime of flavonoids underlying plant-microbe interactions.


Assuntos
Carbono/metabolismo , Flavonoides/metabolismo , Medicago sativa/metabolismo , Microbiologia do Solo , Metais/metabolismo , Minerais/metabolismo , Nitrogênio/metabolismo , Fenômenos Fisiológicos Vegetais , Solo/química
11.
Front Microbiol ; 11: 618373, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33633695

RESUMO

The rapid diversification of synthetic biology tools holds promise in making some classically hard-to-solve environmental problems tractable. Here we review longstanding problems in the Earth and environmental sciences that could be addressed using engineered microbes as micron-scale sensors (biosensors). Biosensors can offer new perspectives on open questions, including understanding microbial behaviors in heterogeneous matrices like soils, sediments, and wastewater systems, tracking cryptic element cycling in the Earth system, and establishing the dynamics of microbe-microbe, microbe-plant, and microbe-material interactions. Before these new tools can reach their potential, however, a suite of biological parts and microbial chassis appropriate for environmental conditions must be developed by the synthetic biology community. This includes diversifying sensing modules to obtain information relevant to environmental questions, creating output signals that allow dynamic reporting from hard-to-image environmental materials, and tuning these sensors so that they reliably function long enough to be useful for environmental studies. Finally, ethical questions related to the use of synthetic biosensors in environmental applications are discussed.

12.
PLoS One ; 13(1): e0191246, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29329343

RESUMO

Biochar may improve soil hydrology by altering soil porosity, density, hydraulic conductivity, and water-holding capacity. These properties are associated with the grain size distributions of both soil and biochar, and therefore may change as biochar weathers. Here we report how freeze-thaw (F-T) cycling impacts the grain size of pine, mesquite, miscanthus, and sewage waste biochars under two drainage conditions: undrained (all biochars) and a gravity-drained experiment (mesquite biochar only). In the undrained experiment plant biochars showed a decrease in median grain size and a change in grain-size distribution consistent with the flaking off of thin layers from the biochar surface. Biochar grain size distribution changed from unimodal to bimodal, with lower peaks and wider distributions. For plant biochars the median grain size decreased by up to 45.8% and the grain aspect ratio increased by up to 22.4% after 20 F-T cycles. F-T cycling did not change the grain size or aspect ratio of sewage waste biochar. We also observed changes in the skeletal density of biochars (maximum increase of 1.3%), envelope density (maximum decrease of 12.2%), and intraporosity (porosity inside particles, maximum increase of 3.2%). In the drained experiment, mesquite biochar exhibited a decrease of median grain size (up to 4.2%) and no change of aspect ratio after 10 F-T cycles. We also document a positive relationship between grain size decrease and initial water content, suggesting that, biochar properties that increase water content, like high intraporosity and pore connectivity large intrapores, and hydrophilicity, combined with undrained conditions and frequent F-T cycles may increase biochar breakdown. The observed changes in biochar particle size and shape can be expected to alter hydrologic properties, and thus may impact both plant growth and the hydrologic cycle.


Assuntos
Carvão Vegetal/química , Solo/química , Ecossistema , Congelamento , Hidrologia , Tamanho da Partícula , Pinus , Poaceae , Porosidade , Prosopis , Esgotos , Temperatura
13.
ACS Synth Biol ; 7(3): 903-911, 2018 03 16.
Artigo em Inglês | MEDLINE | ID: mdl-29366321

RESUMO

Fluorescent proteins are ubiquitous tools that are used to monitor the dynamic functions of natural and synthetic genetic circuits. However, these visual reporters can only be used in transparent settings, a limitation that complicates nondisruptive measurements of gene expression within many matrices, such as soils and sediments. We describe a new ratiometric gas reporting method for nondisruptively monitoring gene expression within hard-to-image environmental matrices. With this approach, C2H4 is continuously synthesized by ethylene forming enzyme to provide information on viable cell number, and CH3Br is conditionally synthesized by placing a methyl halide transferase gene under the control of a conditional promoter. We show that ratiometric gas reporting enables the creation of Escherichia coli biosensors that report on acylhomoserine lactone (AHL) autoinducers used for quorum sensing by Gram-negative bacteria. Using these biosensors, we find that an agricultural soil decreases the bioavailable concentration of a long-chain AHL up to 100-fold. We also demonstrate that these biosensors can be used in soil to nondisruptively monitor AHLs synthesized by Rhizobium leguminosarum and degraded by Bacillus thuringiensis. Finally, we show that this new reporting approach can be used in Shewanella oneidensis, a bacterium that lives in sediments.


Assuntos
Gases/metabolismo , Expressão Gênica , Microbiologia do Solo , Solo , Bacillus thuringiensis/metabolismo , Etilenos/metabolismo , Genes Reporter , Lactonas/metabolismo , Shewanella/metabolismo , Temperatura , Volatilização
14.
Sci Rep ; 7(1): 11233, 2017 09 11.
Artigo em Inglês | MEDLINE | ID: mdl-28894167

RESUMO

Pyrogenic carbon (PyC), produced naturally (wildfire charcoal) and anthropogenically (biochar), is extensively studied due to its importance in several disciplines, including global climate dynamics, agronomy and paleosciences. Charcoal and biochar are commonly used as analogues for each other to infer respective carbon sequestration potentials, production conditions, and environmental roles and fates. The direct comparability of corresponding natural and anthropogenic PyC, however, has never been tested. Here we compared key physicochemical properties (elemental composition, δ13C and PAHs signatures, chemical recalcitrance, density and porosity) and carbon sequestration potentials of PyC materials formed from two identical feedstocks (pine forest floor and wood) under wildfire charring- and slow-pyrolysis conditions. Wildfire charcoals were formed under higher maximum temperatures and oxygen availabilities, but much shorter heating durations than slow-pyrolysis biochars, resulting in differing physicochemical properties. These differences are particularly relevant regarding their respective roles as carbon sinks, as even the wildfire charcoals formed at the highest temperatures had lower carbon sequestration potentials than most slow-pyrolysis biochars. Our results challenge the common notion that natural charcoal and biochar are well suited as proxies for each other, and suggest that biochar's environmental residence time may be underestimated when based on natural charcoal as a proxy, and vice versa.

15.
Environ Sci Technol ; 51(17): 9856-9863, 2017 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-28745499

RESUMO

While it is clear that biochar can alter soil N2O emissions, data on NO impacts are scarce. Reports range from 0 to 67% soil NO emission reductions postbiochar amendment. We use regional air quality and health cost models to assess how these soil NO reductions could influence U.S. air quality and health costs. We find that at 67% soil NO reduction, widespread application of biochar to fertilized agricultural soils could reduce O3 by up to 2.4 ppb and PM2.5 by up to 0.15 µg/m3 in some regions. Modeled biochar-mediated health benefits are up to $4.3 million/county in 2011, with impacts focused in the Midwest and Southwest. These potential air quality and health cobenefits of biochar use highlight the need for an improved understanding of biochar's impacts on soil NO emissions. The benefits reported here should be included with estimates of other biochar benefits, such as crop yield increase, soil water management, and N2O reductions.


Assuntos
Poluentes Atmosféricos , Carvão Vegetal , Óxido Nítrico/análise , Solo , Agricultura , Fertilizantes
16.
PLoS One ; 12(6): e0179079, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28598988

RESUMO

Many studies report that, under some circumstances, amending soil with biochar can improve field capacity and plant-available water. However, little is known about the mechanisms that control these improvements, making it challenging to predict when biochar will improve soil water properties. To develop a conceptual model explaining biochar's effects on soil hydrologic processes, we conducted a series of well constrained laboratory experiments using a sand matrix to test the effects of biochar particle size and porosity on soil water retention curves. We showed that biochar particle size affects soil water storage through changing pore space between particles (interpores) and by adding pores that are part of the biochar (intrapores). We used these experimental results to better understand how biochar intrapores and biochar particle shape control the observed changes in water retention when capillary pressure is the main component of soil water potential. We propose that biochar's intrapores increase water content of biochar-sand mixtures when soils are drier. When biochar-sand mixtures are wetter, biochar particles' elongated shape disrupts the packing of grains in the sandy matrix, increasing the volume between grains (interpores) available for water storage. These results imply that biochars with a high intraporosity and irregular shapes will most effectively increase water storage in coarse soils.


Assuntos
Carvão Vegetal/química , Solo/química , Água , Modelos Teóricos , Tamanho da Partícula , Porosidade
17.
Sci Total Environ ; 592: 316-325, 2017 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-28319718

RESUMO

Although charcoal's density and porosity shape its environmental roles (e.g. controlling its landscape movement and determining the internal pore space available as microbial habitat), the rate of change of these properties in the environment remains largely unknown. With time, charcoal pores may fill or charcoal particles may shatter, altering the ecosystem services delivered. In this study we examined the effects of environmental exposure on the density and porosity of charcoals pyrolyzed at two different temperatures (350 and 520°C). Fresh charcoal made from the dominant local tree (Pinus sylvestris) was returned in litter incubations to Norwegian boreal forest soils for 20months in three different placements: (i) aboveground, (ii) in the humus layer, and (iii) under the humus layer in contact with the mineral subsoil. By varying soil horizon placement we were able to separate the effects of infill from the effects of environmental disturbance on charcoal density and porosity. Environmental exposure changed charcoal density and porosity, and the response varied with environmental placement. Charcoal placed in soil layers increased in porosity by ~1-2% after 20months' incubation. This is likely because open indentations on the charcoal surface became partially occluded, creating more detectable pore space. In contrast, the porosity of charcoal incubated aboveground decreased slightly (~1-2% over 20months). Because there were no minerals or humic substances to infill the aboveground charcoal samples, this porosity reduction was likely caused by breakage of particles induced by weathering. When charcoal particles cleave through pores, internal pore space is destroyed. The small changes observed here indicate that environmental exposure did not trigger rapid shifts in charcoal density and porosity. In addition, these physical properties appear not to have reached equilibrium after 20months incubation, suggesting that the effect of environmental exposure on charcoal's physical properties occurs on the timescale of years to decades.

18.
Environ Sci Technol ; 50(16): 8750-9, 2016 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-27415416

RESUMO

Traditional visual reporters of gene expression have only very limited use in soils because their outputs are challenging to detect through the soil matrix. This severely restricts our ability to study time-dependent microbial gene expression in one of the Earth's largest, most complex habitats. Here we describe an approach to report on dynamic gene expression within a microbial population in a soil under natural water levels (at and below water holding capacity) via production of methyl halides using a methyl halide transferase. As a proof-of-concept application, we couple the expression of this gas reporter to the conjugative transfer of a bacterial plasmid in a soil matrix and show that gas released from the matrix displays a strong correlation with the number of transconjugant bacteria that formed. Gas reporting of gene expression will make possible dynamic studies of natural and engineered microbes within many hard-to-image environmental matrices (soils, sediments, sludge, and biomass) at sample scales exceeding those used for traditional visual reporting.


Assuntos
Solo , Transferases , Biomassa , Genes Microbianos , Microbiologia do Solo
19.
Sci Total Environ ; 563-564: 237-46, 2016 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-27135586

RESUMO

To best use biochar as a sustainable soil management and carbon (C) sequestration technique, we must understand the effect of environmental exposure on its physical and chemical properties because they likely vary with time. These properties play an important role in biochar's environmental behavior and delivery of ecosystem services. We measured biochar before amendment and four years after amendment to a commercial nectarine orchard at rates of 5, 15 and 30tha(-1). We combined two pycnometry techniques to measure skeletal (ρs) and envelope (ρe) density and to estimate the total pore volume of biochar particles. We also examined imbibition, which can provide information about soil hydraulic conductivity. Finally, we investigated the chemical properties, surface, inner layers atomic composition and C1s bonding state of biochar fragments through X-ray photoelectron spectroscopy (XPS). Ageing increased biochar skeletal density and reduced the water imbibition rate within fragments as a consequence of partial pore clogging. However, porosity and the volume of water stored in particles remained unchanged. Exposure reduced biochar pH, EC, and total C, but enhanced total N, nitrate-N, and ammonium-N. X-ray photoelectron spectroscopy analyses showed an increase of O, Si, N, Na, Al, Ca, Mn, and Fe surface (0-5nm) atomic composition (at%) and a reduction of C and K in aged particles, confirming the interactions of biochar with soil inorganic and organic phases. Oxidation of aged biochar fragments occurred mainly in the particle surface, and progressively decreased down to 75nm. Biochar surface chemistry changes included the development of carbonyl and carboxylate functional groups, again mainly on the particle surface. However, changes were noticeable down to 75nm, while no significant changes were measured in the deepest layer, up to 110nm. Results show unequivocal shifts in biochar physical and chemical properties/characteristics over short (~years) timescales.

20.
ACS Omega ; 1(2): 226-233, 2016 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-29938248

RESUMO

The presence of charcoal in soil triggers a range of biological effects that are not yet predictable, in part because it interferes with the functioning of chemical signals that microbes release into their environment to communicate. We do not fully understand the mechanisms by which charcoal alters the biologically available concentrations of these intercellular signals. Recently, charcoal has been shown to sorb the signaling molecules that microbes release, rendering them ineffective for intercellular communication. Here, we investigate a second, potentially more important mechanism of interference: signaling-molecule hydrolysis driven by charcoal-induced soil pH changes. We examined the effects of 10 charcoals on the bioavailable concentration of an acyl-homoserine lactone (AHL) used by many Gram-negative bacteria for cell-cell communication. We show that charcoals decrease the level of bioavailable AHL through sorption and pH-dependent hydrolysis of the lactone ring. We then built a quantitative model that predicts the half-lives of different microbial signaling compounds in the presence of charcoals varying in pH and surface area. Our model results suggest that the chemical effects of charcoal on pH-sensitive bacterial AHL signals will be fundamentally distinct from effects on pH-insensitive fungal signals, potentially leading to shifts in microbial community structures.

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