Biology subjects

Rodriguez-Ramos, J. A.

Publications and source records attributed to Rodriguez-Ramos, J. A..

3 recordsLinked to original sources

bioRxiv2024

Environmental matrix and moisture are key determinants of microbial phenotypes expressed in a reduced complexity soil-analog

Soil moisture and porosity regulate microbial metabolism by influencing factors such as redox conditions, substrate availability, and soil connectivity. However, the inherent biological, chemical, and physical heterogeneity of soil complicates laboratory investigations into microbial phenotypes that mediate community metabolism. This difficulty arises from challenges in accurately representing the soil environment and in establishing a tractable microbial community that limits confounding variables. To address these challenges in our investigation of community metabolism, we use a reduced-complexity microbial consortium grown in a soil analog using a glass-bead matrix amended with chitin. Long-read and short-read metagenomes, metatranscriptomes, metaproteomes, and metabolomes were analyzed to test the effects of soil structure and moisture on chitin degradation. Our soil structure analog system greatly altered microbial expression profiles compared to the liquid-only incubations, emphasizing the importance of incorporating environmental parameters, like pores and surfaces, for understanding microbial phenotypes relevant to soil ecosystems. These changes were mainly driven by differences in overall expression of chitin-degrading Streptomyces species and stress-tolerant Ensifer. Our findings suggest that the success of Ensifer in a structured environment is likely related to its ability to repurpose carbon via the glyoxylate shunt while potentially using polyhydroxyalkanoate granules as a C source. We also identified traits like motility, stress resistance, and biofilm formation that underlie the degradation of chitin across our treatments and inform how they may ultimately alter carbon use efficiency. Together our results demonstrate that community functions like decomposition are sensitive to environmental conditions and more complex than the multi-enzyme pathways involved in depolymerization. ImportanceSoil moisture and porosity are critical mediators of microbial metabolism by influencing factors such as redox conditions, substrate availability, and soil connectivity. However, identifying how microbial community metabolism shifts in response to varying levels of moisture and porosity remains a challenging frontier. This difficulty arises from challenges in accurately representing the soil environment and in establishing tractable microbial communities that limit confounding variables. Moreover, inferring phenotypes based on "key" genes often fails to predict complex phenotypes that arise from cellular interactions. Here, we establish a tractably complex microbial community in a soil analog system amended with chitin and leverage it to understand how microorganisms respond to changes in porosity and moisture. By using genome-resolved metagenomics, metatranscriptomics, and metaproteomics, we report on the microbial lifestyle strategies that underpin changes in community expression like carbon conservation, biofilm production, and stress response.

microbiology↗

bioRxiv2023

Spatial and temporal metagenomics of river compartments reveals viral community dynamics in an urban impacted stream

Although river ecosystems comprise less than 1% of Earths total non-glaciated area, they are critical modulators of microbially and virally orchestrated global biogeochemical cycles. However, most studies either use data that is not spatially resolved or is collected at timepoints that do not reflect the short life cycles of microorganisms. As a result, the relevance of microbiome interactions and the impacts they have over time on biogeochemical cycles are poorly understood. To assess how viral and microbial communities change over time, we sampled surface water and pore water compartments of the wastewater-impacted River Erpe in Germany every 3 hours over a 48-hour period resulting in 32 metagenomes paired to geochemical and metabolite measurements. We reconstructed 6,500 viral and 1,033 microbial genomes and found distinct communities associated with each river compartment. We show that 17% of our vMAGs clustered to viruses from other ecosystems like wastewater treatment plants and rivers. Our results also indicated that 70% of the viral community was persistent in surface waters, whereas only 13% were persistent in the pore waters taken from the hyporheic zone. Finally, we predicted linkages between 73 viral genomes and 38 microbial genomes. These putatively linked hosts included members of the Competibacteraceae, which we suggest are potential contributors to carbon and nitrogen cycling. Together, these findings demonstrate that microbial and viral communities in surface waters of this urban river can exist as stable communities along a flowing river; and raise important considerations for ecosystem models attempting to constrain dynamics of river biogeochemical cycles.

microbiology↗

bioRxiv2022

Genome-resolved metaproteomics decodes the microbial and viral contributions to coupled carbon and nitrogen cycling in river sediments

Rivers have a significant role in global carbon and nitrogen cycles, serving as a nexus for nutrient transport between terrestrial and marine ecosystems. Although rivers have a small global surface area, they contribute substantially to global greenhouse gas emissions through microbially mediated processes within the river hyporheic zone. Despite this importance, microbial roles in these climatically relevant systems are mostly inferred from 16S rRNA amplicon surveys, which are not sufficiently resolved to inform biogeochemical models. To survey the metabolic potential and gene expression underpinning carbon and nitrogen biogeochemical cycling in river sediments, we collected an integrated dataset of over 30 metagenomes, metaproteomes, and paired metabolomes. We reconstructed over 500 microbial metagenome assembled genomes (MAGs), which we dereplicated into 55 unique genomes spanning 12 bacterial and archaeal phyla. We also reconstructed 2482 viral genomic contigs, which were dereplicated into 111 viral MAGs >10kb in size. As a result of integrating gene expression data with geochemical and metabolite data, we created a conceptual model that uncovers new roles for microorganisms in organic matter decomposition, carbon sequestration, nitrogen mineralization, nitrification, and denitrification. Integrated through shared resource pools of ammonium, carbon dioxide, and inorganic nitrogen we show how these metabolic pathways could ultimately contribute to carbon dioxide and nitrous oxide fluxes from hyporheic sediments. Further, by linking viral genomes to these active microbial hosts, we provide some of the first insights into viral modulation of river sediment carbon and nitrogen cycling. ImportanceHere we created HUM-V (Hyporheic Uncultured Microbial and Viral), an annotated microbial and viral genome catalog that captures the strain and functional diversity encoded in river sediments. Demonstrating its utility, this genomic inventory encompasses multiple representatives of the most dominant microbial and archaeal phyla reported in river sediments and provides novel viral genomes that can putatively infect these. Furthermore, we used HUM-V to recruit gene expression data to decipher the functional activities of these genomes and reconstruct their active roles in river sediment biogeochemical cycling. We show the power of genome resolved, multi-omics to uncover the organismal interactions and chemical handoffs shaping an intertwined carbon and nitrogen metabolic network and create a framework that can be extended to other river sediments. The accessible microbial and viral genomes in HUM-V will serve as a community resource to further advance more untargeted, activity-based measurements in these and related freshwater terrestrial-aquatic ecosystems.

microbiology↗