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DeAngelis, K. M.

Publications and source records attributed to DeAngelis, K. M..

11 recordsLinked to original sources

Evidence for endemism and local adaptation in Antarctic soil bacteria

Antarctic soils represent one of the more extreme environments for microbial life on Earth, yet they harbor heterogeneous and diverse microbial communities. Biologists have long hypothesized that Antarctic microorganisms are unique from those found on other continents due to the extreme geographic isolation and the cold, dry, and challenging conditions typical of Antarctica. To test this hypothesis, we focused on a cosmopolitan bacterial genus, Arthrobacter, that is widely distributed across global soils. We first profiled a global metagenomic dataset from both Antarctic and non-Antarctic surface soils to quantify the distributions of Arthrobacter strains. Despite high strain-level diversity, 90% of the strains found in the Antarctic soils were only found on the continent. We then used cultivation-based phenotypic analyses and strain-level genomic comparisons to assess how Antarctic strains and non-Antarctic strains differ in their traits and environmental preferences. Not only did we find evidence of endemism, but Antarctic Arthrobacter also have genomic characteristics and environmental tolerances that suggest they are uniquely adapted to Antarctic conditions. Significance StatementAntarctic soils are among the most extreme environments on Earth, yet they host diverse microbial communities whose adaptations are poorly understood. To test whether Antarctic microbes are distinct from those elsewhere, we examined Arthrobacter, a bacterial genus common in soils worldwide. Analysis of global metagenomic data revealed Arthrobacter strains in Antarctic soils are found exclusively on the continent. Cultivation experiments and comparative genomics further showed that Antarctic strains differ from nonAntarctic relatives in genomic features and environmental tolerances. Together, these results demonstrate that geographically isolated and extreme conditions can drive microbial endemism and local adaptation, even within globally distributed bacterial lineages.

ecology↗

The EPS-I exopolysaccharide transforms Ralstonia wilt pathogen biofilms into viscoelastic fluids for rapid dissemination in planta

Ralstonia solanacearum species complex (RSSC) pathogens cause destructive plant wilt diseases of a wide variety of crops, leading to significant agricultural losses worldwide. These bacteria rapidly spread through the water-transporting xylem where they grow prolifically and produce abundant biofilm that clogs xylem vessels. To understand RSSC biofilm behavior in planta, we examined their complex fluid mechanics. Rheological analyses revealed that unlike all previously analyzed microbial biofilms, RSSC biofilms are shear-thinning, viscoelastic fluids at physiologically relevant shear forces. To determine which factors confer these unique mechanics, we analyzed biofilms of bacterial mutants with altered biofilm components. Genetic analysis demonstrated that development of the viscous-dominant biofilms required production of EPS-I, an amphiphilic exopolysaccharide that is a major virulence factor for all RSSC pathogens. We show that EPS-I confers "biofilm mobility", which allows wildtype RSSC colonies to passively expand when deformed. Despite its high metabolic cost, bioassays demonstrated that EPS-I production conferred a net fitness benefit where biofilm mobility allowed the pathogen to spread and access more nutrients in complex environments like xylem vessels. The RSSC are a monophyletic lineage of aggressive plant wilt pathogens, and our evolutionary hypothesis testing suggests the origin of the eps biosynthetic gene cluster coincides with the emergence of wilt pathogenesis in the RSSC ancestor. Furthermore, comparative physiological assays demonstrated that biofilm mobility is unique to the RSSC within the genus Ralstonia. In summary, EPS-I production is a key evolutionary innovation that enables RSSC dispersal and virulence by conferring unique biofilm mechanics. Significance StatementRalstonia solanacearum species complex (RSSC) pathogens threaten global food security by fatally wilting plants. A soft matter physics lens demystified the cryptic role of a major virulence factor, the EPS-I exopolysaccharide. EPS-I transforms RSSC biofilms into viscoelastic fluids, a mechanical behavior not previously described for other microbial biofilms that are almost always viscoelastic solids. We demonstrate that the development of fluid biofilms was a key evolutionary innovation that enabled pathogenic success of these aggressive pathogens that rapidly wilt plants.

microbiology↗

Long-term warming inverts the relationship between ecosystem function and microbial resource acquisition

Soil microbial traits drive ecosystem functions, which can explain the positive correlation between microbial functional diversity and ecosystem function. However, microbial adaptation to climate change related warming stress can shift microbial traits with direct implications for soil carbon cycling. Here, we investigated how long-term warming affects the relationship between microbial trait diversity and ecosystem function. Soils were sampled after 24 years of +5{degrees}C warming alongside unheated control soils from the Harvard Forest Long-Term Ecological Research site. Ecosystem function was estimated from six different enzyme activities and microbial biomass. Functional diversity was calculated from metatranscriptomics sequencing, where reads were assigned to yield, acquisition, or stress trait categories. We found that in organic horizon soils, warming decreased the richness of acquisition-related traits. In the mineral soils, we observed that heated soils exhibited a negative relationship with the richness of acquisition-related traits. These results suggest that microbial communities exposed to long-term warming are shifting away from a resource acquisition life history strategy.

ecology↗

Season, not long-term warming, affects the relationship between ecosystem function and microbial diversity

1Across biomes, soil biodiversity promotes ecosystem functions. However, whether this relationship will be maintained under climate change is uncertain. Here, using two long-term soil warming experiments, we investigated how warming affects the relationship between ecosystem functions and bacterial diversity across seasons, soil horizons, and warming duration. Soils were sampled from these warming experiments located at the Harvard Forest Long-Term Ecological Research (LTER) site, where soils had been heated +5 {degrees}C above ambient for 13 or 28 years at the time of sampling. We assessed seven measurements representative of different ecosystem functions and nutrient pools. We also surveyed bacterial community diversity. We found that ecosystem function was significantly affected by season, with autumn samples having higher function than summer samples. The effect of warming on bacterial diversity was similarly affected by season, where warming in the summer was associated with decreased bacterial evenness in the organic horizon. Despite the decreased bacterial diversity in the warmed plots, we found that the relationship between ecosystem function and bacterial diversity was unaffected by warming or warming duration. Our findings highlight that season is a consistent driver of ecosystem function as well as a modulator of climate change effects on bacterial community diversity.

ecology↗

Thermal adaptation of soil microbial growth traits in response to chronic warming

Adaptation of soil microbes due to warming from climate change has been observed, but it remains unknown what microbial growth traits are adaptive to warming. We studied bacterial isolates from the Harvard Forest Long-Term Ecological Research site, where field soils have been experimentally heated to 5{degrees}C above ambient temperature with unheated controls for thirty years. We hypothesized that Alphaproteobacteria from warmed plots have (1) less temperature sensitive growth rates; (2) higher optimum growth temperatures; and (3) higher maximum growth temperatures compared to isolates from control plots. We made high-throughput measurements of bacterial growth in liquid cultures over time and across temperatures from 22-37{degrees}C in 2-3{degrees}C increments. We estimated growth rates by fitting Gompertz models to the growth data. Temperature sensitivity of growth rate, optimum growth temperature, and maximum growth temperature were estimated by the Ratkowsky 1983 model and a modified Macromolecular Rate Theory (MMRT) model. To determine evidence of adaptation, we ran phylogenetic generalized least squares tests on isolates from warmed and control soils. Our results showed evidence of adaptation of higher optimum growth temperature of bacterial isolates from heated soils. However, we observed no evidence of adaptation of temperature sensitivity of growth and maximum growth temperature. Our project begins to capture the shape of the temperature response curves, but illustrates that the relationship between growth and temperature is complex and cannot be limited to a single point in the biokinetic range. ImportanceSoils are the largest terrestrial carbon sink and the foundation of our food, fiber, and fuel systems. Healthy soils are carbon sinks, storing more carbon than they release. This reduces the amount of carbon dioxide released to the atmosphere and buffers against climate change. Soil microbes drive biogeochemical cycling and contribute to soil health through organic matter breakdown, plant growth promotion, and nutrient distribution. In this study, we determined how soil microbial growth traits respond to long-term soil warming. We found that bacterial isolates from warmed plots showed evidence of adaptation of increased optimum growth temperature. This suggests that increased microbial biomass and growth relative to respiration in a warming world should result in greater carbon storage. As temperatures increase, greater microbial activity may help reduce the soil carbon feedback loop. Our results provide insight on how atmospheric carbon cycling and soil health may respond in a warming world.

microbiology↗

Pangenomes reveal genomic signatures of microbial adaptation to chronic soil warming

Below-ground carbon transformations represent a natural climate change mitigation solution, but newly-acquired traits adaptive to climate stress may alter microbial climate feedback mechanisms. To better define microbial evolutionary responses to long-term climate warming, we study microorganisms from an ongoing in situ soil warming experiment at the Harvard Forest Long-term Ecological Research (LTER) site where, for over three decades, soils are continuously heated 5 {degrees}C above ambient temperatures. We hypothesize that across generations of chronic warming, genomic signatures within diverse bacterial lineages reflect trait-based adaptations related to growth and carbon utilization. From our bacterial culture collection isolated from experimental heated and control plots, we sequenced genomes representing taxa dominant in soil communities and sensitive to warming, including lineages of Alphaproteobacteria, Actinobacteria, and Betaproteobacteria. We investigated differences in genomic attributes and patterns of functional gene content to identify genomic signatures of adaptation. Comparative pangenomics revealed accessory gene clusters related to central metabolism, competition, and carbon substrate degradation. Overall, genomes from control plots were relatively enriched in carbon and fatty acid metabolism pathways, while genomes from heated plots were relatively enriched in nitrogen metabolism pathways. We also observed differences in global codon usage bias between heated and control genomes, suggesting potential adaptive traits related to growth or growth efficiency. This effect was more varied for organisms with fewer 16S rrn operons, suggesting that these organisms experience different selective pressures on growth efficiency. Together, these data illustrate the emergence of lineage-specific traits as well as common ecological-evolutionary microbial responses to climate change.

microbiology↗

Substrate availability and not thermal-acclimation controls microbial temperature sensitivity response to long term warming

Microbes are responsible for cycling carbon (C) through soils, and the predictions of how soil C stocks change with warming are highly sensitive to the assumptions made about the mechanisms controlling the microbial physiology response to climate warming. Two mechanisms, microbial thermal-acclimation and changes in the quantity and quality of substrates available for microbial metabolism have been suggested to explain the long-term warming impact on microbial physiology. Yet studies disentangling these two mechanisms are lacking. To resolve the drivers of changes in microbial physiology in response to long-term warming, we sampled soils from 13- and 28-year old soil warming experiments in different seasons. We performed short-term laboratory incubations across a range of temperatures to measure the relationship between temperature sensitivity of physiology (growth, respiration, carbon use efficiency and extracellular enzyme activity) and the chemical composition of soil organic matter. We observed apparent thermal acclimation in microbial processes important for C cycling, but only when warming had exacerbated the seasonally-induced, already small soil organic matter pools. Irrespective of warming, greater quantity and quality of soil carbon enhanced the extracellular enzymatic pool and its temperature sensitivity. We suggest that fresh litter input into the system seasonally cancels apparent thermal acclimation of C-cycling processes. Our findings reveal that long-term warming has indirectly affected microbial physiology via reduced C availability in this system, implying that earth system models including these negative feedbacks may be best suited to describe long-term warming impact in soils.

microbiology↗

Chitosan diet alters the microbiome of adult house flies

House flies are disease vectors, carrying human pathogens which include Escherichia coli and Vibrio cholera. To explore the use of chitosan as a bioinsecticide, we evaluated the effects of a chitosan-amended diet on Musca domestica (house fly). We first conducted longevity experiments to understand the impact of chitosan on house fly longevity. We confirmed that chitosan diet amendment is associated with reduced longevity and that this is not due to starvation. We then extracted fly microbiome DNA and used 16S ribosomal RNA gene amplicon sequencing and quantitative PCR to assess the composition and load of the microbiome for flies fed chitosan-amended diets compared to controls. Diversity of the chitosan-fed fly microbiomes was lower than the control, with significant dissimilarities in community composition. Chitosan-fed flies showed lower Ralstonia relative abundance but increased relative abundance of Serratia. Both control and chitosan-fed flies had highly uneven communities, but the control flies were dominated by genera Ralstonia and Providencia, while the chitosan-fed flies were dominated by genera Serratia, Kosakonia, and Providencia. Contrary to our expected results, chitosan-fed flies also contained 56% more bacteria compared to controls. Gut microbiome changes appear to result from chitinolytic bacteria becoming more relatively abundant, and our results suggest that chitosan-amended diet alters the house fly microbiome resulting in higher fly mortality.

microbiology↗

The role of microbial diversity in the formation of soil organic matter quality and persistence

The largest terrestrial carbon sink on earth is soil carbon stocks. As the climate changes, the rate at which the Earths climate warms depends in part on the persistence of soil organic carbon. Microbial turnover forms the backbone of soil organic matter (SOM) formation and it has been recently proposed that SOM molecular complexity is a key driver of stability. Despite this, the links between microbial diversity, chemical complexity and biogeochemical nature of soil organic matter remain missing. Here we used a model soil system to test the hypothesis that more diverse microbial communities generate more stable soil organic matter. We inoculated microbial communities of varying diversities into an model soil matrix amended with simple carbon, and measured the thermal stability of the resultant soil organic matter. Using a novel data analysis approach with Rock-Eval(R) ramped thermal analysis, we found that microbial community diversity drives the chemical fingerprint of soil organic matter. Bacteria-only and low diversity communities lead to less chemically-diverse and more thermally-labile soil carbon pools than highly diverse communities. Our results provide direct evidence for a link between microbial diversity, molecular complexity and SOM stability. This evidence demonstrates the benefits of managing soils for maximum biological diversity as a means of building persistent SOM stocks. ClassificationBiological Sciences: Ecology

ecology↗

Iron Chelator-Mediated Anoxic Biotransformation of Lignin by Novel sp., Tolumonas lignolytica BRL6-1

Lignin is the second most abundant carbon polymer on earth and despite having more fuel value than cellulose, it currently is considered a waste byproduct in many industrial lignocellulose applications. Valorization of lignin relies on effective and green methods of delignification, with a growing interest in the use of microbes. Here we investigate the physiology and lignin biotransformation mechanisms of the novel facultative anaerobic bacterium, Tolumonas lignolytica BRL6-1, under anoxic conditions. Physiological and biochemical changes were compared between cells grown anaerobically in either lignin-amended or unamended conditions. In the presence of lignin, BRL6-1 had a higher biomass and shorter lag phase compared to unamended conditions, and 14% of the proteins determined to be significantly higher in abundance by log2 fold-change of 2 or greater were related to Fe(II) transport in early exponential phase. Ferrozine assays of the supernatant (<10 kDa fraction) confirmed that Fe(III) was bound to lignin and reduced to Fe(II) only in the presence of BRL6-1, suggesting redox activity by the cells. LC-MS/MS analysis of the secretome showed an extra band at 20 kDa in lignin-amended conditions. Protein sequencing of this band identified a protein of unknown function with homology to enzymes in the radical SAM superfamily. Expression of this protein in lignin-amended conditions suggests its role in radical formation. From our findings, we suggest that BRL6-1 is using a protein in the radical SAM superfamily to interact with the Fe(III) bound to lignin and reducing it to Fe(II) for cellular use, increasing BRL6-1 yield under lignin-amended conditions. This interaction potentially generates organic free radicals and causes a radical cascade which could modify and depolymerize lignin. Further research should clarify the extent to which this mechanism is similar to previously described aerobic chelator-mediated Fenton chemistry or radical producing lignolytic enzymes, such as lignin peroxidases, but under anoxic conditions.

microbiology↗

Heavy and wet: evaluating the validity and implications of assumptions made when measuring growth efficiency using 18O water

1How microbes allocate carbon to growth vs. respiration plays a central role in determining the ability of soil to retain carbon. This carbon use efficiency (CUE) is increasingly measured using the 18O-H2O method, in which heavy oxygen incorporated into DNA is used to estimate growth. Here we evaluated the validity of some of the assumptions of this method using a literature search, and then tested how violating them affected estimates of the growth component of carbon use efficiency in soil. We found that the 18O method is consistently sensitive to assumptions made about oxygen sources to DNA, but that the effect of other assumptions depends on the microbial community present. We provide an example for how the tools developed here may be used with observed CUE values, and demonstrate that the original conclusions drawn from the data remain robust in the face of methodological bias. Our results lay the foundation for a better understanding of the consequences to the 18O method underlying assumptions. Future studies can use the approach developed here to identify how different incubation conditions and/or treatments might bias its CUE estimates and how trustworthy their results are. Further wet-lab work dissecting the assumptions of the 18O method in soil will help justify the scenarios under which it is reasonable to trust its results.

microbiology↗