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At least 55 records · Page 3

Time-lapse approach to correct deficiencies of 2D soil zymography

Membrane zymography is commonly used in rhizosphere ecology to map enzyme activities in intact soil samples and plant roots. The method consists of incubating a membrane saturated with an enzyme-specific fluorogenic substrate on the soil/root surface followed by measurements of fluorescence intensity of the product in the membrane. The traditional zymography is based on assumptions that fluorescence on membrane images is linearly increasing with time during zymography and an increase in product content is numerically equal to enzyme activity in soil below the membrane. These assumptions are unlikely to hold in experimental settings. In this work we introduce a new zymography technique, time-lapse zymography (TLZ); the approach that eliminates the need for assumptions of the traditional zymography and provides more realistic estimates of enzymatic activities. We assessed the new technique in a series of laboratory and modeling experiments, including quantification of the fluorescent product diffusion (e.g. MUF: 4-methylumbelliferone) from plant roots, enzyme activity measurements on the roots, and HYDRUS-2D & HP2 software calibration with obtained data. The calibrated model was used to analyze the processes governing spatial and temporal dynamics of MUF contents in the membrane. The results indicated that the enzyme diffusion within the membrane-soil system was negligible, and measured zymograms were adequately reproduced solely by accounting for substrate and product diffusions and for catalytic enzyme reaction described by the Michaelis-Menten equation. TLZ enabled identifying and using linear parts on MUF time series and accounting for MUF losses in each zymogram pixel, considerably improving the accuracy as compared to the traditional method. Results demonstrated that enzymatic activity from only a thin soil layer (~0.2 mm) is adequately represented in zymograms.

54 ENVIRONMENTAL SCIENCES↗

Community RNA-Seq: multi-kingdom responses to living versus decaying roots in soil

Abstract Roots are a primary source of organic carbon input in most soils. The consumption of living and detrital root inputs involves multi-trophic processes and multiple kingdoms of microbial life, but typical microbial ecology studies focus on only one or two major lineages. We used Illumina shotgun RNA sequencing to conduct PCR-independent SSU rRNA community analysis (“community RNA-Seq”) and simultaneously assess the bacteria, archaea, fungi, and microfauna surrounding both living and decomposing roots of the annual grass, Avena fatua. Plants were grown in 13CO2-labeled microcosms amended with 15N-root litter to identify the preferences of rhizosphere organisms for root exudates (13C) versus decaying root biomass (15N) using NanoSIMS microarray imaging (Chip-SIP). When litter was available, rhizosphere and bulk soil had significantly more Amoebozoa, which are potentially important yet often overlooked top-down drivers of detritusphere community dynamics and nutrient cycling. Bulk soil containing litter was depleted in Actinobacteria but had significantly more Bacteroidetes and Proteobacteria. While Actinobacteria were abundant in the rhizosphere, Chip-SIP showed Actinobacteria preferentially incorporated litter relative to root exudates, indicating this group’s more prominent role in detritus elemental cycling in the rhizosphere. Our results emphasize that decomposition is a multi-trophic process involving complex interactions, and our methodology can be used to track the trajectory of carbon through multi-kingdom soil food webs.

Nuccio, Erin E. (ORCID:000000030189183X)↗

The rhizosphere and cropping system, but not arbuscular mycorrhizae, affect ammonia oxidizing archaea and bacteria abundances in two agricultural soils

Arbuscular mycorrhizal fungi (AMF) form symbioses with roots that can enhance plant nutrition. While AMF have been shown to have a role in soil nitrogen (N) cycling, it is unclear whether AMF affect N cycling microbes such as ammonia-oxidizing bacteria (AOB) and archaea (AOA), which convert ammonium into nitrite in the first step of nitrification. In this study, we examined the effects of AMF on AOA and AOB abundances within the corn rhizosphere and bulk soil of conventional (corn-soybean rotation with inorganic fertilizer) and diversified (cornsoybean- oats/alfalfa-oats rotation with composted manure) systems. We hypothesized that AMF would decrease AOA and AOB abundances in a cropping-system dependent manner, possibly due to competition for ammonium. We grew corn deficient or proficient in AMF symbiosis in microcosms for 10 weeks. At the end of the experiment, both soils planted with the AMF-proficient corn genotype had higher ammonium and lower nitrate pool sizes compared to the same soils planted with the AMF-deficient corn genotype. Likewise, total plant N was higher in the AMF-proficient genotype compared to the AMF-deficient genotype. Despite changes in soil inorganic N pool sizes, AOA and AOB abundances were unaffected by plant AMF-proficiency. Instead, AOA abundance was greater in the rhizosphere than in the bulk soil regardless of cropping system, and AOB abundance was greater in the conventional than the diversified cropping system soil regardless of proximity to the root. These data indicate that 1) AMF did not affect AOA or AOB abundance in these N-rich soils but other factors such as root proximity and inorganic fertilization did and 2) AOA and AOB have differing ecological niches within rhizosphere and bulk soil that should be considered when managing for nitrogen losses.

Wattenburger, Cassandra↗

Dual-mycorrhizal colonization is determined by plant age and host identity in two species of Populus

Plants have evolved symbioses with mycorrhizal and endophytic fungi that are essential for their growth and survival. While most plants associate with a single guild of mycorrhizal fungi, a select group termed “dual-mycorrhizal plants” associate with both arbuscular mycorrhizal and ectomycorrhizal fungi. Although a shift from predominance of arbuscular mycorrhizal to ectomycorrhizal colonization with plant development has been demonstrated on other dual-mycorrhizal hosts, it is not known how mycorrhizal colonization shifts with plant age in Populus species. We performed a controlled growth experiment with natural field-sourced inocula to test for age-dependent shifts in fungal colonization rates and for host-specific patterns of colonization in two species of Populus (P. tremuloides and P. trichocarpa). We found that only P. trichocarpa displayed dual-mycorrhizal colonization, while P. tremuloides associated with ectomycorrhizal fungi, but not arbuscular mycorrhizal fungi. Both guilds of mycorrhizal fungi increased in abundance with plant age, while root endophytic fungal colonization decreased. Many of the early-colonizing endophytic fungi that we documented have strong saprotrophic capabilities, which may be an important trait for fast colonization. Dark septate endophytes were more abundant than either guild of mycorrhizal fungi, and are likely to be functionally important members of the Populus root fungal community. Our findings represent a novel pattern in the development of dual-mycorrhizal colonization and illustrate that Populus species vary in their association with arbuscular mycorrhizal fungi. In conclusion, our results also highlight the importance of dark septate endophyte colonization dynamics on dual-mycorrhizal plants.

59 BASIC BIOLOGICAL SCIENCES↗

Vulnerability of mineral-organic associations in the rhizosphere

The majority of soil carbon (C) is stored in organic matter associated with reactive minerals. These mineral-organic associations (MOAs) inhibit microbial and enzymatic access to organic matter, suggesting that organic C within MOAs is resistant to decomposition. However, plant roots and rhizosphere microbes are known to transform minerals through dissolution and exchange reactions, implying that MOAs in the rhizosphere can be dynamic. Here we identify key drivers, mechanisms, and controls of MOA disruption in the rhizosphere and present a new conceptual framework for the vulnerability of soil C within MOAs. We introduce a vulnerability spectrum that highlights how MOAs characteristic of certain ecosystems are particularly susceptible to specific root-driven disruption mechanisms. This vulnerability spectrum provides a framework for critically assessing the importance of MOA disruption mechanisms at the ecosystem scale. Comprehensive representation of not only root-driven MOA formation, but also disruption, will improve model projections of soil C-climate feedbacks and guide the development of more effective soil C management strategies.

54 ENVIRONMENTAL SCIENCES↗

The Largely Ignored but Important Role of Non‐Rooted, Submersed Vegetation and Small Waterbodies on Greenhouse Gas Emissions

Abstract Small, shallow waterbodies are abundant globally and are associated with greenhouse gas (GHG) emissions disproportionate to their size. One reason for this is the prevalence of non‐rooted, submersed (NRS) macrophytes that often dominate these systems. Despite these plants' significant role in the carbon ecology of their ecosystems, they are often excluded from climate change models due, in part, to the lack of clarity of their effects. Theus et al. (2023, https://doi.org/10.1029/2023JG007758 ) provides new insights into the role that NRS macrophytes have on GHG emissions, demonstrating that increasing densities of these macrophytes lead to increases in both CO 2 and CH 4 gas fluxes. Here, we highlight the importance of both shallow waterbodies and NRS vegetation within the context of the field of aquatic ecosystem biogeochemistry. We identify a need for the inclusion of these drivers into climate change models based partly on the findings of Theus et al. (2023, https://doi.org/10.1029/2023JG007758 ).

Hassett, Erin↗

metagRoot: a comprehensive database of protein families associated with plant root microbiomes

The plant root microbiome is vital in plant health, nutrient uptake, and environmental resilience. To explore and harness this diversity, we present metagRoot, a specialized and enriched database focused on the protein families of the plant root microbiome. MetagRoot integrates metagenomic, metatranscriptomic, and reference genome-derived protein data to characterize 71 091 enriched protein families, each containing at least 100 sequences. These families are annotated with multiple sequence alignments, CRISPR elements, hidden Markov models, taxonomic and functional classifications, ecosystem and geolocation metadata, and predicted 3D structures using AlphaFold2. MetagRoot is a powerful tool for decoding the molecular landscape of root-associated microbial communities and advancing microbiome-informed agricultural practices by enriching protein family information with ecological and structural context. The database is available at https://pavlopoulos-lab.org/metagroot/ or https://www.metagroot.org.

Chasapi, Maria N↗

Burning questions for a warming and changing world: 15 unknowns in plant abiotic stress

Abstract We present unresolved questions in plant abiotic stress biology as posed by 15 research groups with expertise spanning eco-physiology to cell and molecular biology. Common themes of these questions include the need to better understand how plants detect water availability, temperature, salinity, and rising carbon dioxide (CO2) levels; how environmental signals interface with endogenous signaling and development (e.g. circadian clock and flowering time); and how this integrated signaling controls downstream responses (e.g. stomatal regulation, proline metabolism, and growth versus defense balance). The plasma membrane comes up frequently as a site of key signaling and transport events (e.g. mechanosensing and lipid-derived signaling, aquaporins). Adaptation to water extremes and rising CO2 affects hydraulic architecture and transpiration, as well as root and shoot growth and morphology, in ways not fully understood. Environmental adaptation involves tradeoffs that limit ecological distribution and crop resilience in the face of changing and increasingly unpredictable environments. Exploration of plant diversity within and among species can help us know which of these tradeoffs represent fundamental limits and which ones can be circumvented by bringing new trait combinations together. Better defining what constitutes beneficial stress resistance in different contexts and making connections between genes and phenotypes, and between laboratory and field observations, are overarching challenges.

Biochemistry & Molecular Biology↗

Tundra Vegetation Community Type, Not Microclimate, Controls Asynchrony of Above‐ and Below‐Ground Phenology

The below-ground growing season often extends beyond the above-ground growing season in tundra ecosystems and as the climate warms, shifts in growing seasons are expected. However, we do not yet know to what extent, when and where asynchrony in above- and below-ground phenology occurs and whether variation is driven by local vegetation communities or spatial variation in microclimate. Here, we combined above- and below-ground plant phenology metrics to compare the relative timings and magnitudes of leaf and fine-root growth and senescence across microclimates and plant communities at five sites across the Arctic and alpine tundra biome. We observed asynchronous growth between above- and below-ground plant tissue, with the below-ground season extending up to 74% (~56 days) beyond the onset of above-ground leaf senescence. Plant community type, rather than microclimate, was a key factor controlling the timing, productivity, and growth rates of fine roots, with graminoid roots exhibiting a distinct ‘pulse’ of growth later into the growing season than shrub roots. Our findings indicate the potential of vegetation change to influence below-ground carbon storage as the climate warms and roots remain active in unfrozen soils for longer. Taken together, our findings of increased root growth in soils that remain thawed later into the growing season, in combination with ongoing tundra vegetation change including increased shrub and graminoid abundance, indicate increased below-ground productivity and altered carbon cycling in the tundra biome.

below-ground↗

Drought shifts dissolved organic matter sources from above- to belowground and stress-induced processes in Amazon white-sand forests

White-sand forests contribute significantly to dissolved organic matter (DOM) production in the central Amazon, forming blackwater rivers that dominate organic matter export from the Amazon basin to the ocean. Despite their importance in controlling DOM export, white-sand forests are understudied, and it remains unclear whether systematic changes in the formation of blackwater DOM occur and how seasonal variations and extremes like El Niño-associated droughts impact them. We collected soil porewater from two central Amazon white-sand forests for two years, spanning a wet La Niña year followed by an El Niño drought year. The molecular composition of DOM was analyzed using high-resolution mass spectrometry, and correlation network analysis was employed to identify ecologically meaningful DOM subsets. Using additional chemical characterization, database annotations, correlation with 14C-age of DOM and climatic variables, and ecological null modeling, we propose five distinct DOM sources: plant litter and throughfall, soil organic matter (SOM) decomposition, root exudation, and two drought response subsets of likely microbial and plant origin. During drought conditions, aboveground plant-derived compounds decreased, while SOM products, root exudates, and drought response compounds increased. These drought responses were qualitatively similar in both years but notably amplified in the drier El Niño year. Drought amplified deterministic control over DOM composition, indicating that DOM reflected directed biological responses and that future droughts are likely to generate similar shifts. Overall, drought substantially altered belowground carbon cycling by shifting DOM sources and inducing stress responses, effects expected to recur and potentially intensify under future climate scenarios.

Lange, Dan F.↗

The Role of Synthetic Biology in Atmospheric Greenhouse Gas Reduction: Prospects and Challenges

The long atmospheric residence time of CO 2 creates an urgent need to add atmospheric carbon drawdown to CO 2 regulatory strategies. Synthetic and systems biology (SSB), which enables manipulation of cellular phenotypes, offers a powerful approach to amplifying and adding new possibilities to current land management practices aimed at reducing atmospheric carbon. The participants (in attendance: Christina Agapakis, George Annas, Adam Arkin, George Church, Robert Cook-Deegan, Charles DeLisi, Dan Drell, Sheldon Glashow, Steve Hamburg, Henry Jacoby, Henry Kelly, Mark Kon, Todd Kuiken, Mary Lidstrom, Mike MacCracken, June Medford, Jerry Melillo, Ron Milo, Pilar Ossorio, Ari Patrinos, Keith Paustian, Kristala Jones Prather, Kent Redford, David Resnik, John Reilly, Richard J. Roberts, Daniel Segre, Susan Solomon, Elizabeth Strychalski, Chris Voigt, Dominic Woolf, Stan Wullschleger, and Xiaohan Yang) identified a range of possibilities by which SSB might help reduce greenhouse gas concentrations and which might also contribute to environmental sustainability and adaptation. These include, among other possibilities, engineering plants to convert CO 2 produced by respiration into a stable carbonate, designing plants with an increased root-to-shoot ratio, and creating plants with the ability to self-fertilize. A number of serious ecological and societal challenges must, however, be confronted and resolved before any such application can be fully assessed, realized, and deployed.

59 BASIC BIOLOGICAL SCIENCES↗

A Test of Functional Balance Theory for Wetland Biomass Allocation in a Global Change Experiment

Abstract Forecasts of root growth and carbon sequestration under global change are compromised by uncertainty in how plants will allocate biomass between above and belowground pools. Here, we develop a simple model to assess whether functional balance theory can explain a complex biomass allocation response observed in a brackish marsh under experimental warming and elevated CO 2 . Our model shows how treatment‐driven changes in nitrogen supply and demand can explain divergent observations of root growth (i.e., maximum responses under intermediate warming and elevated CO 2 ). The model also reveals a surprising interaction between warming and eutrophication, where enhanced N loading to coastal marshes may reduce adverse impacts of warming on root growth. Our findings provide a mechanistic basis for incorporating biomass allocation into forecast models of marsh evolution. They also provide a general example of using ecological theory to decompose complex net responses observed in multi‐factor global change experiments into constituent processes.

54 ENVIRONMENTAL SCIENCES↗

Fine Root and Soil Organic Carbon Depth Distributions are Inversely Related Across Fertility and Rainfall Gradients in Lowland Tropical Forests

Humid tropical forests contain some of the largest soil organic carbon (SOC) stocks on Earth. Much of this SOC is in subsoil, yet variation in the distribution of SOC through the soil profile remains poorly characterized across tropical forests. We used a correlative approach to quantify relationships among depth distributions of SOC, fine root biomass, nutrients and texture to 1 m depths across 43 lowland tropical forests in Panama. The sites span rainfall and soil fertility gradients, and these are largely uncorrelated for these sites. We used fitted b parameters to characterize depth distributions, where b is a numerical index based on an asymptotic relationship, such that larger b values indicate greater concentrations of root biomass or SOC at depth in the profile. Root b values ranged from 0.82 to 0.95 and were best predicted by soil pH and extractable potassium (K) stocks. For example, the three most acidic (pH < 4) and Kpoor (< 20 g K m -2 ) soils contained 76 ± 5% of fine root biomass from 0 to 10 cm depth, while the three least acidic (pH > 6.0) and most K-rich (> 50 g K m -2 ) soils contained only 41 ± 9% of fine root biomass at this depth. Root b and SOC b values were inversely related, such that a large fine root biomass in surface soils corresponded to large SOC stocks in subsoils (50–100 cm). SOC b values were best predicted by soil pH and base cation stocks, with the three most base-poor soils containing 34 ± 8% of SOC from 50 to 100 cm depth, and the three most base-rich soils containing just 9 ± 2% of SOC at this depth. Nutrient depth distributions were not related to Root b or SOC b values. Here, these data show that large surface root biomass stocks are associated with large subsoil C stocks in strongly weathered tropical soils. Further studies are required to evaluate why this occurs, and whether changes in surface root biomass, as may occur with global change, could in turn influence SOC storage in tropical forest subsoils.

54 ENVIRONMENTAL SCIENCES↗

Chronic drought differentially alters the belowground microbiome of drought tolerant and drought susceptible genotypes of Populus trichocarpa

Populus trichocarpa is an ecologically important tree species and economically important biofeedstock. Belowground, P. trichocarpa interacts with diverse microorganisms in the rhizosphere and root endosphere. These plant-microbial interactions can bolster a variety of plant processes, ranging from pathogen suppression to drought tolerance, yet we know little about the impact of chronic drought stress on P. trichocarpa’s belowground microbiomes. To investigate the interactive effect of chronic drought on belowground microbial communities across genetically different P. trichocarpa hosts, we assessed archaeal/bacterial and fungal communities within the root endosphere, rhizosphere, and surrounding bulk soil of selected genotypes in a long-term drought experiment in Boardman, OR, USA. We sequenced the 16S rRNA and ITS2 gene region on samples collected from 16 distinct P. trichocarpa genotypes in plots with full or reduced irrigation. Eight of these genotypes have been previously identified as drought tolerant while the other eight genotypes were drought susceptible. While reduced irrigation influenced the composition of every archaeal/bacterial microbiome compartment, fungal communities were only affected in the rhizosphere and bulk soil compartments. Drought-tolerant bacteria, such as Actinobacteria, were differentially abundant in reduced irrigation across all belowground microbiomes. Host drought-tolerance influenced plant-associated microbiome compartments but had little impact on the bulk soil compartment. Drought-tolerant trees were enriched for potential growth-promoting microorganisms in the root endosphere and rhizosphere, including Sphingomonas bacteria and ectomycorrhizal fungi. Overall, associations of growth-promoting microbes in drought resistant P. trichocarpa genotypes can be leveraged to improve biofeedstock productivity in regions prone to periodic drought.

59 BASIC BIOLOGICAL SCIENCES↗

Widespread co‐occurrence of Sebacinales and arbuscular mycorrhizal fungi in switchgrass roots and soils has limited dependence on soil carbon or nutrients

Societal Impact Statement This work addresses a novel group of Sebacinales mycorrhizal fungi being considered for development as inoculants in managed ecosystems because of their potential access to nutrients in soil organic matter. By comparing the diversity, distributions, and abundances of the Sebacinales with the more common arbuscular mycorrhizal fungi in switchgrass, a biofuel crop, we demonstrate that current suggestions for a Sebacinales revolution in agriculture should be tempered by their ecology. In particular, Sebacinales are rare compared to arbuscular mycorrhizal fungi, and are only weakly associated with soil carbon, suggesting that ideas about improved soil carbon cycling associated with Sebacinales need to be sufficiently studied across a range of environmental conditions prior to their consideration for broad‐spectrum soil inoculants. Summary Arbuscular mycorrhizal (AM) fungi are widespread and important root symbionts, but recent work suggests that Sebacinales fungi may play an equally important role in both plant success and ecosystem carbon and nutrient cycling based on their worldwide occurrence and putative access to organic matter. However, the ecological impacts of Sebacinales will depend on their abundance and distribution relative to AM fungi and environmental soil carbon and nutrient patterns, which remain unexplored. We characterized Glomeromycota and Sebacinales fungi in switchgrass ( Panicum virgatum L.) roots and soils across 14 sites with diverse soil conditions. We examined group richness differences, co‐occurrence patterns, and how the relative abundance of these fungi related to soil carbon, nutrient stoichiometry, and host size. Sebacinales were widespread, but less diverse, common, and abundant than Glomeromycota. Moreover, co‐occurrences were predominantly random, suggesting relatively few interactions between these groups. Sebacinales increased relative to Glomeromycota in soils with more carbon, but explanatory power was limited. Based on our findings, we suggest that Sebacinales are likely complementary to AM fungi in roots. Expectations that Sebacinales have large effects on soil carbon and nutrient cycling may need to be reconsidered, at least based on their limited abundances relative to AM fungi in switchgrass. This is an important consideration as Sebacinales are candidates for use as inoculants in managed ecosystems.

Lee, Marissa R.↗

Aligning theoretical and empirical representations of soil carbon-to-nitrogen stoichiometry with process-based terrestrial biogeochemistry models

Soil carbon-nitrogen (C:N) stoichiometry acts as a control over decomposition and soil organic matter formation and loss, making it a key soil property for understanding ecosystem dynamics and projected ecosystems responses to global environmental change. However, the controls of soil C:N and how they respond to increasing pressures from global change agents are not fully understood. The “foundational” controls on soil C:N, namely plant and microbial C:N, have been used to predict soil C:N, but fail to accurately simulate all ecosystems and may be insufficient for predictions under global environmental change. Here we present an “emerging” representation of controls of soil C:N that includes plant-microbe-mineral feedbacks that have been shown to regulate soil C:N. We argue that including representation of these emerging drivers in process-based terrestrial biogeochemistry models, which include biological N fixation, mycorrhizae, priming, root exudation of organic acids, and mineralogy (including soil texture, mineral composition, and aggregation), will improve mechanistic representation of soil C:N and associated processes. Such improvements will produce models that will better simulate a variety of ecological states and predict soil C:N when global changes modify plant-microbe-mineral interactions. Here, we align our empirical understanding of controls of soil C:N with those controls represented in models, identifying contexts where emerging drivers might be particularly important to represent (e.g., priming and root exudation in nutrient-limited conditions) and areas of future work. Additionally, we show that implementing emerging drivers of soil C:N results in different simulated outcomes at steady state and in response to elevated atmospheric CO2. Our review and preliminary simulations support the need to incorporate emerging drivers of soil C:N into process-based terrestrial biogeochemistry models, allowing for both theoretical exploration of mechanisms and potentially more accurate predictions of land biogeochemical responses to global change.

54 ENVIRONMENTAL SCIENCES↗