Biology subjects

Yee, M. O.

Publications and source records attributed to Yee, M. O..

6 recordsLinked to original sources

bioRxiv2023

Decoding Root Biogeography: Building Reduced Complexity Functional Rhizosphere Microbial Consortia

The rhizosphere microbiome plays a crucial role in supporting plant productivity and contributes to ecosystem functioning by regulating nutrient cycling, soil integrity, and carbon storage. However, characterizing their functional attributes and microbial relationships remains challenging due to their complex taxonomic and functional compositions. To enable such studies, the development of reduced complexity microbial consortia derived from the rhizosphere microbiome of the natural ecosystem is highly desirable. Designing and assembling reduced complexity consortia that mimic natural communities with consistent, stable, predictable features are highly sought after but is challenging to deliver. Here we present our systematic controlled design towards successful assembly of several such rhizosphere derived reduced complexity consortia. From Brachypodium grown in natural soil under controlled lab conditions, we enriched the root-associated microbes, utilizing carbon compounds prevalent in Brachypodium root exudates. By transferring the enrichments every 3 or 7 days for 9 generations, we developed both fast and slow-growing microbial communities. 16S rRNA amplicon analysis revealed that both inoculum and carbon substrates significantly influence microbial community composition. For example, 1/10 R2A preferentially enriched Amplicon Sequence Variants (ASVs) from slow growing taxa vital to plant including Acidobacteria and Verrucomicrobia. Network analysis revealed that although fast and slow growing microbial consortia have distinct key taxa, the key hubs (keystone taxa) for both belong to genera with plant growth promoting (PGP) traits. This suggests that PGP bacteria might play a central role in controlling the microbial networks among rhizospheric microbiomes. Based on the stability and richness results from different transfers, most carbon substrates lead to microbial consortia with reduced complexity and high stability after a few transfers. The stability tests of the derived microbial consortia also showed high stability, reproducibility, and revivability of the constructed microbial consortia. Our study represents a significant step towards understanding and harnessing the potential of rhizosphere microbiomes, with implications for sustainable agriculture and environmental management.

microbiology↗

bioRxiv2023

Fine scale sampling reveals spatial heterogeneity of rhizosphere microbiome in young Brachypodium plants

For a deeper and comprehensive understanding of the diversity, composition and function of rhizosphere microbiomes, we need to focus at the scale of individual roots in standardized growth containers. Root exudation patterns are known to vary across distinct parts of the root giving rise to spatially distinct microbial niches. To address this, we analyzed microbial community from two spatially distinct zones of the primary root (the tip vs. the base) in Brachypodium distachyon, grown in natural soil using standardized fabricated ecosystems known as EcoFABs as well as in more conventional pot and tubes. 16S rRNA based community analysis showed a stronger rhizosphere effect in the root base vs. bulk soil compared to the root tips vs. bulk soil, resulting in an enrichment of Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria, few OTUs belonging to less characterized lineages such as Verrucomicrobia and Acidobacteria. While the microbial community distributions are similar across growth containers, the EcoFAB displayed higher replicate reproducibility. Genome-resolved and bulk metagenomics revealed that genes associated with transcriptional regulation, transport of nutrients and catabolic enzymes indicating active metabolism, biofilm formation and root colonization were enriched in root tips. On the other hand, genes associated with nutrient-limitation and environmental stress were prominent in the bulk soil compared to the root tips, implying the presence of easily available, labile carbon and nutrients in the rhizosphere relative to bulk soil. Such insights into the relationships between root structure, exudation and microbial communities are critical for developing understanding of plant-microbe interactions.

ecology↗

bioRxiv2022

The Fate of DIET Consortium exposed to continuous and intermittent electrochemical stimulation

Electromethanogenesis is the electrochemical conversion of carbon dioxide (CO2) to methane (CH4) with microorganisms as catalysts, and it is a promising approach for biogas upgrading. Many studies have shown increased methane production by electrochemical stimulation, allegedly due to the enhancement of direct interspecies electron transfer (DIET) between electrogenic bacteria and methanogens. However, these studies were done with mixed cultures where the interspecies interactions were undefined, so the impact of an electrochemical stimulus on these interactions remains unresolved. Here we report that an electrochemical stimulus diminished the survival and activity of a defined DIET consortium. We investigated the impact of electrochemical stimulation on a co-culture of Geobacter metallireducens and Methanosarcina barkeri, where the two partners interact syntrophically via DIET. The electrochemical stimulus was a cathode poised at - 700 mV (versus the standard hydrogen electrode, SHE). Electrochemical stimulation was provided either continuously or intermittently, and so was the food substrate. Compared to the untreated DIET co-culture, all treatment strategies relying on cathodic current additions resulted in acetate accumulation, lower methane production and lower cell numbers for both partners, indicating a detrimental impact of electrochemical stimulation to a DIET consortium. These findings suggest that future bioelectrochemical technologies must guarantee the survival of the syntrophic partners during current additions.

microbiology↗

bioRxiv2020

Culturing of "Unculturable" Subsurface Microbes: Natural Organic Carbon Source Fuels the Growth of Diverse and Distinct Bacteria from Groundwater

The recovery and cultivation of diverse field-related microorganisms from the terrestrial subsurface environment remains a challenge despite recent advances in modern molecular technology. Here we applied natural organic carbon (C), i.e., sediment-derived natural organic matter (NOM) and bacterial cell lysate, to groundwater microbial communities for a 30-day enrichment incubation, followed by conventional direct-plating for isolation. The groundwater was collected from a background well at the Oak Ridge Reservation Field Research Center, Tennessee. As a comparison, we also included enrichments amended with simple organic C sources, including glucose, acetate, benzoate, oleic acid, cellulose, and mixed vitamins. Our results demonstrate that complex natural organic C sources are more effective in enriching diverse bacterial species from groundwater than simple organic C sources. Microcosms amended with simple organic C (glucose, acetate, benzoate, or oleic acid) show significantly lower biodiversity than unamended control and are dominated by only few phyla such as Proteobacteria and Bacteroidetes. In contrast, microcosms amended with complex natural organic C (sediment NOM or bacterial cell lysate) display significantly higher biodiversity, and enrich distinct species from the phyla that are poorly represented in published culture collections (e.g., Verrucomicrobia, Planctomycetes, and Armatimonadetes). Our subsequent isolation efforts from natural organic C-amended enrichments led to 222 purified bacterial isolates representing 5 phyla, 16 orders, and 54 distinct species including candidate novel, rarely cultivated, and undescribed organisms. ImportanceInnovative strategies for recovering bacterial strains representing the true diversity of microbial communities in the terrestrial subsurface would significantly advance the understanding of ecologically critical taxa residing in these ecosystems. In this study, we demonstrate that complex natural organic C that mimic the naturally available resources for microbes encourages the growth of diverse bacteria much more robustly than traditional simplistic organic C sources. Results from this study will substantially advance and improve the design of strategies to effectively cultivate and isolate diverse and novel subsurface microorganisms in the laboratory. Obtaining axenic cultures of the once-unculturable microorganisms will greatly enhance our understanding of microbial physiology, function, and roles in different biogeochemical niches in terrestrial subsurface ecosystems.

microbiology↗

bioRxiv2019

Extracellular electron uptake in Methanosarcinales is independent of multiheme c-type cytochromes

The co-occurrence of Geobacter and Methanosarcinales is often used as a proxy for the manifestation of direct interspecies electron transfer (DIET) in man-made and natural aquatic environments. We previously reported that not all Geobacter are capable of DIET with Methanosarcina. Here we tested 15 new artificial co-culture combinations with methanogens and electrogenic bacteria, including an electrogen outside of the Geobacter clade - Rhodoferax ferrireducens.\n\nConsistently, highly effective electrogenic bacteria (G. metallireducens, G. hydrogenophilus and R. ferrireducens) formed successful associations with Methanosarcinales. Highly effective electrogens could not sustain the growth of H2-utilizing methanogens of the genera Methanococcus, Methanobacterium, Methanospirillum, Methanolacinia or Methanoculleus.\n\nMethanosarcinales, including strict non-hydrogenotrophic methanogens of the genus Methanothrix (Mtx. harundinacea and Mtx. shoeghenii) and Methanosarcina horonobensis, conserved their ability to interact with electrogens. Methanosarcinales were classified as the only methanogens containing c-type cytochromes, unlike strict hydrogenotrophic methanogens. It was then hypothesized that multiheme c-type cytochromes give Methanosarcinales their ability to retrieve extracellular electrons. However, multiheme c-type cytochromes are neither unique to this group of methanogens nor universal. Only two of the seven Methanosarcinales tested had multiheme c-type cytochromes (MCH). In one of these two species - M. mazei a deletion mutant for its MCH was readily available. Here we tested if the absence of this MHC impacts extracellular electron uptake. Deletion of the MHC in M. mazei did not impact the ability of this methanogens to retrieve extracellular electrons from G. metallireducens or a poised cathode. Since Methanosarcina did not require multiheme c-type cytochromes for direct electron uptake we proposed an alternative strategy for extracellular electron uptake.

microbiology↗

bioRxiv2018

Extracellular electron uptake by two Methanosarcina species

Direct electron uptake by prokaryotes is a recently described mechanism with a potential application for energy and CO2 storage into value added chemicals. Members of Methanosarcinales, an environmentally and biotechnologically relevant group of methanogens, were previously shown to retrieve electrons from an extracellular electrogenic partner performing Direct Interspecies Electron Transfer (DIET) and were therefore proposed to be electroactive. However, their intrinsic electroactivity has never been examined. In this study, we tested two methanogens belonging to the genus Methanosarcina, M. barkeri and M. horonobensis, regarding their ability to accept electrons directly from insoluble electron donors like other cells, conductive particles and electrodes. Both methanogens were able to retrieve electrons from Geobacter metallireducens via DIET. Furthermore, DIET was also stimulated upon addition of electrically conductive granular activated carbon (GAC) when each was co-cultured with G. metallireducens. However, when provided with a cathode poised at -400 mV (vs. SHE), only M. barkeri could perform electromethanogenesis. In contrast, the strict hydrogenotrophic methanogen, Methanobacterium formicicum, did not produce methane regardless of the type of insoluble electron donor provided (Geobacter cells, GAC or electrodes). A comparison of functional gene categories between the two Methanosarcina showed differences regarding energy metabolism, which could explain dissimilarities concerning electromethanogenesis at fixed potentials. We suggest that these dissimilarities are minimized in the presence of an electrogenic DIET partner (e.g. Geobacter), which can modulate its surface redox potentials by adjusting the expression of electroactive surface proteins.

microbiology↗