Karrie A. Weber

A hydrogenotrophic methanogen, designated strain ACI-7 ^T , was isolated from an alkaline, saline wetland located in eastern Nebraska, USA. Cells were identified as non-motile rods (0.9–4.2 µm in length and 0.2–0.4 µm in diameter) that occurred singly, in chains, or as twisted chains and aggregates. Strain ACI-7 ^T utilized H ₂ +CO ₂ or formate as methanogenic substrates and required growth factors present in yeast extract for continuous cultivation. The strain grew at 20–45 °C (optimum, 40 °C), at pH 6.5–8.5 (optimum, pH 7.3) and with 0–2.5% NaCl (optimum, 0–1%). The genomic G+C content of ACI-7 ^T was 31.79 mol%. Phylogenetic analysis of the 16S rRNA gene sequence indicated strain ACI-7 ^T was affiliated with the genus Methanobacterium , most closely related to Methanobacterium oryzae FPi ^T (97.1% sequence similarity) and Methanobacterium veterum MK4 ^T (96.9% sequence similarity). Overall genome relatedness indices for strain ACI-7 ^T compared to other Methanobacterium species ranged from 68.45 to 78.17% for average nucleotide identity and 18.9 to 25.7% for digital DNA–DNA hybridization. Morphological, physiological and genomic characteristics indicate that strain ACI-7 ^T represents a novel species, for which the name Methanobacterium nebraskense sp. nov. is proposed. The type strain is ACI-7 ^T (=DSM 118696 ^T =ATCC TSD-487 ^T ).

A hydrogenotrophic methanogen, designated strain ACI-7 ^T , was isolated from an alkaline, saline wetland located in eastern Nebraska, USA. Cells were identified as non-motile rods (0.9–4.2 μm in length and 0.2–0.4 µm in diameter) that occurred singly, in chains, and as long filaments and twisted aggregates. Strain ACI-7 ^T utilized H ₂ + CO ₂ or formate as methanogenic substrates and required growth factors present in yeast extract for continuous cultivation. The strain grew at 20–45°C (optimum, 40°C), at pH 6.5–8.5 (optimum, pH 7.3) and with 0–2.5% NaCl (optimum, 0–1%). The genomic G+C content of ACI-7 ^T was 31.79 mol%. Phylogenetic analysis of the 16S rRNA gene sequence indicated strain ACI-7 ^T was affiliated with the genus Methanobacterium, most closely related to Methanobacterium oryzae FPi ^T (97.1% sequence similarity) and Methanobacterium veterum MK4 ^T (96.9% sequence similarity). Overall genome relatedness indices for strain ACI-7 ^T compared to other Methanobacterium species ranged from 68.45–78.17% for average nucleotide identity and 18.9–25.7% for digital DNA–DNA hybridization. Morphological, physiological, and genomic characteristics indicate that strain ACI-7 ^T represents a novel species, for which the name Methanobacterium nebraskense sp. nov. is proposed. The type strain is ACI-7 ^T (=DSM 118696 ^T =ATCC TSD-487 ^T ).

Methanobacterium sp. ACI-7 was isolated from an enrichment culture established with calcium carbonate as the sole inorganic carbon source originating from alkaline–saline wetland soil. The genome sequence consisted of a single, circular 2,239,949 bp chromosome. A total of 2,366 protein-coding genes, 37 tRNAs, and 6 rRNAs were predicted.

Carbonate minerals are considered a metastable carbon reservoir under alkaline conditions. Here we demonstrate the use of calcium carbonate as a sole inorganic carbon source at moderately alkaline pH for a methanogenic microbial community enriched from alkaline, saline wetland soil. In reactors amended with calcium carbonate as the sole source of inorganic carbon, concentrations of methane and aqueous calcium increased concurrent with headspace hydrogen depletion. Cells were observed in association with the carbonate mineral matrix via coherent anti-Stokes Raman scattering and two-photon excitation fluorescence microscopy. Genome-resolved metagenomics of the enrichment community confirmed the presence of a methanogen (Methanobacterium sp.) with the metabolic potential for hydrogenotrophic methanogenesis in addition to five bacterial community members. These results demonstrate that carbonate minerals can serve as an inorganic carbon source for hydrogenotrophic methanogenesis, leading to mineral dissolution under stable alkaline conditions. As such, hydrogenotrophic methanogens may impact carbon biogeochemistry and carbonate mineral stability in environmental systems.

Plants influence the abiotic and biotic environment of the rhizosphere, affecting plant performance through plant–soil feedback (PSF). We compared the strength of nutrient and microbe-mediated PSF and its implications for plant performance in domesticated and wild grasses with a fully crossed greenhouse PSF experiment using four inbred maize genotypes (Zea mays ssp. mays b58, B73-wt, B73-rth3, and HP301), teosinte (Z. mays ssp. parviglumis), and two wild prairie grasses (Andropogon gerardii and Tripsacum dactyloides) to condition soils for three feedback species (maize B73-wt, teosinte, Andropogon gerardii). We found evidence of negative PSF based on growth, phenotypic traits, and foliar nutrient concentrations for maize B73-wt, which grew slower in maize-conditioned soil than prairie grass-conditioned soil. In contrast, teosinte and A. gerardii showed few consistent feedback responses. Both rhizobiome and nutrient-mediated mechanisms were implicated in PSF. Based on 16S rRNA gene amplicon sequencing, the rhizosphere bacterial community composition differed significantly after conditioning by prairie grass and maize plants, and the final soil nutrients were significantly influenced by conditioning, more so than by the feedback plants. These results suggest PSF-mediated soil domestication in agricultural settings can develop quickly and reduce crop productivity mediated by PSF involving changes to both the soil rhizobiomes and nutrient availability.

Plant genotype is recognized to contribute to variations in microbial community structure in the rhizosphere, soil adherent to roots. However, the extent to which the viral community varies has remained poorly understood and has the potential to contribute to variation in soil microbial communities. Here we cultivated replicates of two Zea mays genotypes, parviglumis and B73, in a greenhouse and harvested the rhizobiome (rhizoplane and rhizosphere) to identify the abundance of cells and viruses as well as rhizobiome microbial and viral community using 16S rRNA gene amplicon sequencing and genome resolved metagenomics. Our results demonstrated that viruses exceeded microbial abundance in the rhizobiome of parviglumis and B73 with a significant variation in both the microbial and viral community between the two genotypes. Of the viral contigs identified only 4.5% (n = 7) of total viral contigs were shared between the two genotypes, demonstrating that plants even at the level of genotype can significantly alter the surrounding soil viral community. An auxiliary metabolic gene associated with glycoside hydrolase (GH5) degradation was identified in one viral metagenome-assembled genome (vOTU) identified in the B73 rhizobiome infecting Propionibacteriaceae (Actinobacteriota) further demonstrating the viral contribution in metabolic potential for carbohydrate degradation and carbon cycling in the rhizosphere. This variation demonstrates the potential of plant genotype to contribute to microbial and viral heterogeneity in soil systems and harbors genes capable of contributing to carbon cycling in the rhizosphere.

Mobilization of naturally-occurring uranium(U) has been recognized to give rise to geogenic U groundwater contamination in aquifers. In addition to carbonate ligand complexation, nitrate has been demonstrated to play a role in controlling U mobility by altering uranium solubility through redox reactions. Nitrate is a common anthropogenic contaminant often prevalent at high concentrations in alluvial aquifers overlaying managed land. Alluvial deposition processes that form these aquifers create a lithologically heterogeneous subsurface with defined contacts between sands, silts, and clays. This leads to deposition of organic carbon and accumulation of reduced metals/radionuclides, including U(IV), in the finer grained silts and clays. The addition of high nitrate porewater into uranium-bearing alluvial aquifer silt sediments stimulated a nitrate reducing microbial community capable of catalyzing U(IV) oxidation and mobilization of U into porewaters. However, metadata from an aquifer wide study and a subsequent experiment revealed that this result is concentration dependent. Low concentrations of nitrate bearing pore-water added into organic-rich, uranium bearing sediments and resulted in a decrease in dissolved U(VI), consistent with reduction. XANES analysis of sediments supported U(VI) reduction with the precipitation of U(IV). U(VI) reduction activity occurred concurrent with an increase in dissolved organic carbon (DOC) and cell and virus abundance and activity. Metagenome assembled genomes from the microbial community revealed the metabolic potential indicating complex carbon degradation, fermentation, mineralization as well as the potential for anaerobic respiration of nitrate, metal/radionuclides, and sulfate. The virome recovered from the samples indicated a change in viral community in response to nitrate amendments and viral-encoded carbohydrate active enzymes were upregulated indicating a coupled response of both viral and microbial community regulating nitrate stimulated carbon biogeochemical cycling. These data together suggest that the addition of an electron acceptor in to organic carbon reduced sediments stimulates not only microbial but also viral activity leading to upregulation of genes associated with carbon biogeochemical cycling in sedimentary systems. While genes associated with metal oxidation are observed, net reduction of uranium prevails leading to uranium immobilization at low nitrate concentrations. Thus together these data indicate a tipping point whereby the influx of nitrate into the reduced environment can influence uranium mobility in DOC and carbon cycling supporting microbial activity and reducing conditions subsurface systems.

Plant genotype is recognized to contribute to variations in microbial community structure in the rhizosphere, soil adherent to roots. However, the extent to which the viral community varies has remained poorly understood and has the potential to contribute to variation in soil microbial communities. Here we cultivated replicates of two different genotypes of Zea mays parviglumis and Z. mays genotype B73 in a greenhouse and harvested the rhizobiome (rhizoplane and rhizosphere) to identify the abundance of cells and viruses as well as apply 16S rRNA gene amplicon sequencing and genome resolved metagenomics to identify the rhizobiome microbial and viral community. Our results demonstrate that viruses exceeded microbial abundance in the rhizobiome of parviglumis and B73 with a significant variation in both, the microbial and viral community between the two genotypes. Of the viral contigs identified only 4.5% (n =7) of total viral contigs were shared between the two genotypes, demonstrating that plants even at the level of genotype can significantly alter the surrounding soil viral community. An auxiliary metabolic gene associated with glycoside hydrolase (GH5) degradation was identified in one viral metagenome-assembled genome (vMAG) identified in the B73 rhizobiome infecting Propionibacteriaceae (Actinobacteriota) further demonstrating the viral contribution in metabolic potential for carbohydrate degradation and carbon cycling in the rhizosphere. This variation demonstrates the potential of plant genotype to contribute to microbial and viral heterogeneity in soil systems and harbor genes capable of contributing to carbon cycling in the rhizosphere.

Groundwater uranium (U) concentrations have been measured above the U.S. EPA maximum contaminant level (30 μg/L) in many U.S. aquifers, including in areas not associated with anthropogenic contamination by milling or mining. In addition to carbonate, nitrate has been correlated to uranium groundwater concentrations in two major U.S. aquifers. However, to date, direct evidence that nitrate mobilizes naturally occurring U from aquifer sediments has not been presented. Here, we demonstrate that the influx of high-nitrate porewater through High Plains alluvial aquifer silt sediments bearing naturally occurring U(IV) can stimulate a nitrate-reducing microbial community capable of catalyzing the oxidation and mobilization of U into the porewater. Microbial reduction of nitrate yielded nitrite, a reactive intermediate, which was further demonstrated to abiotically mobilize U from the reduced alluvial aquifer sediments. These results indicate that microbial activity, specifically nitrate reduction to nitrite, is one mechanism driving U mobilization from aquifer sediments in addition to previously described bicarbonate-driven desorption from mineral surfaces, such as Fe(III) oxides.