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At least 1,405 records · Page 78

Deriving pattern from complexity in the processes underlying tropical forest drought impacts

Tropical forests play a disproportionately large role in the regulation of the Earth’s climate due to their influence over the carbon, water, and energy cycles (e.g. Pan et al., 2011). Questions exist regarding the future contribution of tropical forests to climate regulation, however, due to uncertainties surrounding both chronic and discrete changes in the environment and their subsequent effects on the terrestrial system (Friedlingstein et al., 2006; Friend et al., 2014). Among the most spatially and temporally pervasive threats to the structure and function of tropical forests are those of stronger, more frequent droughts, atmospheric warming, and their combination (Davidson et al., 2012). There is already evidence of these changes. Two ‘100-year’ droughts occurred in theAmazon in 2005 and 2010 (Lewis et al., 2011), and climate models consistently suggest that the length of the dry season will increase (reviewed by McDowell et al., in this issue of New Phytologist, pp. 851–869). Simultaneously, the atmospheric demand for water (vapor pressure deficit; VPD) increases with temperature (Trenberth et al., 2014), placing greater stress on the hydraulic supply capacity of plants (McDowell & Allen, 2015). Finally, a lengthening of the dry season and the warming of the atmosphere are superimposed over the El Ni~no Southern Oscillation (ENSO) pattern – such as the 2015–2016 ENSO– which creates drier-thannormal conditions throughout much of the tropics (IPCC, 2014). Thus, there is a confluence of large predictive uncertainty with the high-importance of the future of the tropical carbon cycle.

McDowell, Nate G.↗

Complex interactions between day length and diurnal patterns of gene expression drive photoperiodic responses in a perennial C 4 grass

Abstract Photoperiod is a key environmental cue affecting flowering and biomass traits in plants. Key components of the photoperiodic flowering pathway have been identified in many species, but surprisingly few studies have globally examined the diurnal rhythm of gene expression with changes in day length. Using a cost‐effective 3′‐Tag RNA sequencing strategy, we characterize 9,010 photoperiod responsive genes with strict statistical testing across a diurnal time series in the C 4 perennial grass, Panicum hallii . We show that the vast majority of photoperiod responses are driven by complex interactions between day length and sampling periods. A fine‐scale contrast analysis at each sampling time revealed a detailed picture of the temporal reprogramming of cis ‐regulatory elements and biological processes under short‐ and long‐day conditions. Phase shift analysis reveals quantitative variation among genes with photoperiod‐dependent diurnal patterns. In addition, we identify three photoperiod enriched transcription factor families with key genes involved in photoperiod flowering regulatory networks. Finally, coexpression networks analysis of GIGANTEA homolog predicted 1,668 potential coincidence partners, including five well‐known GI‐interacting proteins. Our results not only provide a resource for understanding the mechanisms of photoperiod regulation in perennial grasses but also lay a foundation to increase biomass yield in biofuel crops.

Weng, Xiaoyu↗

Phytosulfokine downregulates defense‐related WRKY transcription factors and attenuates pathogen‐associated molecular pattern‐triggered immunity

SUMMARY Phytosulfokine (PSK) is a plant growth‐promoting peptide hormone that is perceived by its cell surface receptors PSKR1 and PSKR2 in Arabidopsis. Plants lacking the PSK receptors show phenotypes consistent with PSK signaling repressing some plant defenses. To gain further insight into the PSK signaling mechanism, comprehensive transcriptional profiling of Arabidopsis treated with PSK was performed, and the effects of PSK treatment on plant defense readouts were monitored. Our study indicates that PSK's major effect is to downregulate defense‐related genes; it has a more modest effect on the induction of growth‐related genes. WRKY transcription factors (TFs) emerged as key regulators of PSK‐responsive genes, sharing commonality with a pathogen‐associated molecular pattern (PAMP) responses, flagellin 22 (flg22), but exhibiting opposite regulatory directions. These PSK‐induced transcriptional changes were accompanied by biochemical and physiological changes that reduced PAMP responses, notably mitogen‐activated protein kinase (MPK) phosphorylation (previously implicated in WRKY activation) and the cell wall modification of callose deposition. Comparison with previous studies using other growth stimuli (the sulfated plant peptide containing sulfated tyrosine [PSY] and Pseudomonas simiae strain WCS417) also reveals WRKY TFs' overrepresentations in these pathways, suggesting a possible shared mechanism involving WRKY TFs for plant growth–defense trade‐off.

Liu, Dian [Biochemistry and Molecular Biophysics T↗

Solids Erosion Patterns Developed by Pulse Jet Mixers

Millions of gallons of radioactive waste are stored in underground storage tanks at the Hanford Site in Washington State. This waste will be vitrified at the Waste Treatment and Immobilization Plant that is under construction. Vessels in the pretreatment portion of the plant are being configured for processing waste slurries with challenging physical and rheological properties that range from Newtonian slurries to non-Newtonian sludge. Pulse jet mixing technology has been selected for mobilizing and mixing this waste. In the pulse jet mixing process, slurry is expelled from pulse tube nozzles directed towards the vessel floor. The expelled fluid forms a radial jet that erodes the settled layer of solids. The pulse tubes are configured in a ring or multiple rings and operate concurrently. The expelled fluid and mobilized solids traverse toward the center of the tank. At the tank center the jets from pulse tubes in the ring collide and lift solids upward in a central plume. At the end of the pulse, when the desired fluid volume is expelled from the pulse tube, the applied pressure switches to suction and the pulse tube is refilled. This cycle is used to mobilize and mix the tank contents. An initial step of the process is the erosion of solids from the vessel floor by the radial jets that form on the vessel flow beneath each pulse tube. Experiments have been conducted using simulants to evaluate the ability of the pulse jet mixing system radial jets to combine to develop the central upwell and lift solids into the vessel. These experiments have been conducted at three scales using a range of granular simulants over a range of concentrations. The vessels have elliptical, spherical, or flanged and dished bottoms. Process parameters evaluated include the velocity of fluid expelled from the pulse tube, the duration of the pulse and the duty cycle, the ratio of pulse duration to cycle time. Videos taken from beneath the vessel show the growth of the cleared area from each pulse tube as a function of time. All solids are lifted from the vessel bottom when the system is operating at the critical suspension velocity. The focus of this paper is to compare and contrast erosion patterns developed from different simulants and pulse tube configurations. The cases are evaluated to determine how changes in process parameters affects the PJM ability to mobilize solids from the vessel floor.

erosion↗

Machine-learning-enabled on-the-fly analysis of RHEED patterns during thin film deposition by molecular beam epitaxy

Thin film deposition is a fundamental technology for the discovery, optimization, and manufacturing of functional materials. Deposition by molecular beam epitaxy (MBE) typically employs reflection high-energy electron diffraction (RHEED) as a real-time in situ probe of the growing film. However, the state-of-the-art for RHEED analysis during deposition requires human observation. Here, we present an approach using machine learning (ML) methods to monitor, analyze, and interpret RHEED images on-the-fly during thin film deposition. In the analysis workflow, RHEED pattern images are collected at one frame per second and featurized using a pretrained deep convolutional neural network. The feature vectors are then statistically analyzed to identify changepoints; these changepoints can be related to changes in the deposition mode from initial film nucleation to a transition regime, smooth film deposition, and in some cases, an additional transition to a rough, islanded deposition regime. The feature vectors are additionally analyzed via graph analysis and community classification. The graph is quantified as a stabilization plot, and we show that inflection points in the stabilization plot correspond to changes in the growth regime. The full RHEED analysis workflow is termed RHAAPsody and includes data transfer and output to a visual dashboard. We demonstrate the functionality of RHAAPsody by analyzing the precaptured RHEED images from epitaxial depositions of anatase TiO2 on SrTiO3(001) and show that the analysis workflow can be executed in less than 1 s. Our approach shows promise as one component of ML-enabled real-time feedback control of the MBE deposition process.

36 MATERIALS SCIENCE↗

Orientation control of high-χ triblock copolymer for sub-10 nm patterning using fluorine-containing polymeric additives

Directed self-assembly (DSA) of block copolymers (BCPs) Is one of the most promising techniques to tackle the ever-increasing demand for subllthographic features In semiconductor industries. BCPs with high Flory-Huggins parameter (chi) are of particular Interest due to their ability to self-assemble at the length scale of sub-10 nm. However, such high-chi BCPs typically have imbalanced surface energies between respective blocks, making it a challenge to achieve desired perpendicular orientation. To address this challenge, we mixed a fluorine-containing polymeric additive with poly(2-vlnylpyrldine)-block-polystyrene-block-poly(2-vlnylpyrldine) (P2VP-b-PS-b-P2VP) and successfully controlled the orientation of the high-chi triblock copolymer. The additive selectively mixes with P2VP block through hydrogen bonding and can reduce the dissimilarity of surface energies between PS and P2VP blocks. After optimizing additive dose and annealing conditions, desired perpendicular orientation formed upon simple thermal annealing. We further demonstrated DSA of this material system with five times density multiplication and a half-pitch as small as 8.5 nm. This material system Is also amenable to sequential Infiltration synthesis treatment to selectively grow metal oxide In P2VP domains, which can facilitate the subsequent pattern transfer. We believe that this integration-friendly DSA platform using simple thermal annealing holds the great potential for sub-10 nm nanopatterning applications.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Patterns of patient specific dosimetry in total body irradiation

Purpose: Total body irradiation (TBI) has been used for bone marrow transplant for hematologic and immune deficiency conditions. The goal of TBI is to deliver a homogeneous dose to the entire body, with a generally accepted range of dose uniformity being within {+-}10% of the prescribed dose. The moving table technique for TBI could make dose uniform in whole body by adjusting couch speed. However, it is difficult to accurately estimate the actual dose by calculation and hence in vivo dosimetry (IVD) is routinely performed. Here, the authors present patterns of patient-specific IVD in 161 TBI patients treated at our institution. Methods: Cobalt-60 teletherapy unit (Model C9 Cobalt-60 teletherapy unit, Picker X-ray Corporation) with customized moving bed (SITI Industrial Products, Inc., Fishers, IN) were used for TBI treatment. During treatment, OneDose{sup TM} (Sicel Technology, NC) Metal Oxide-silicon Semiconductor Field Effect Transistor detectors were placed at patient body surface; both entrance and exit side of the beam at patient head, neck, mediastinum, umbilicus, and knee to estimate midplane dose. When large differences (>10%) between the prescribed and measured dose were observed, dose delivery was corrected for subsequent fractions by the adjustment of couch speed and/or bolus placement. Under IRB exempt status, the authors retrospectively analyzed the treatment records of 161 patients who received TBI treatment between 2006 and 2011. Results: Across the entire cohort, the median {+-} SD (range) percent variance between calculated and measured dose for head, neck, mediastinum, umbilicus, and knee was -2.3 {+-} 10.2% (-66.2 to +35.3), 1.1 {+-} 11.5% (-62.2 to +40.3), -1.9 {+-} 9.5% (-66.4 to +46.6), -1.1 {+-} 7.2% (-35.2 to +42.9), and 3.4 {+-} 12.2% (-47.9 to +108.5), respectively. More than half of treatments were within {+-}10% of the prescribed dose for all anatomical regions. For 80% of treatments (10%-90%), dose at the umbilicus was within {+-}10%. However, some large differences greater than 35% were also found at several points. For one case, the knee received double the prescribed dose. When the dose differences for multiple fractions were averaged, compliance ({+-}10%) between the prescription and measured dose was improved compared to the dose difference of the first single fraction, for example, as at umbilicus, which improved from 83.9% to 98.5%. Conclusions: Actual dose measurement analysis of TBI patients revealed a potentially wide variance from the calculated dose. Based from their IVD method for TBI using Cobalt-60 irradiator and moving table, {+-}10% over entire body is hard to achieve. However, it can be significantly improved with immediate feedback after the first fraction prior to subsequent treatments.

60 APPLIED LIFE SCIENCES↗

TU-C-17A-05: Dose Domain Optimization of MLC Leaf Patterns for Highly Complicated 4Ï€ IMRT Plans

Purpose: Highly conformal non-coplanar 4π radiotherapy plans typically require more than 20 intensity-modulated fields to deliver. A novel method to calculate multileaf collimator (MLC) leaf patterns is introduced to maximize delivery efficiency, accuracy and plan quality. Methods: 4 GBM patients, with a prescription dose of 59.4 Gy or 60 Gy, were evaluated using the 4π algorithm using 20 beams. The MLC calculation utilized a least square minimization of the dose distribution, with an anisotropic total variation regularization term to encourage piecewise continuity in the fluence maps. Transforming the fluence to the dose domain required multiplying the fluence with a sparse matrix. Exploiting this property made it feasible to solve the problem using CVX, a MATLAB-based convex modeling framework. The fluence was stratified into even step sizes, and the MLC segments, limited to 300, were calculated. The patients studied were replanned using Eclipse with the same beam angles. Results: Compared to the original 4π plan, the stratified 4π plan increased the maximum/mean dose for, in Gy, by 1.0/0.0 (brainstem), 0.5/0.2 (chiasm), 0.0/0.0 (spinal cord), 1.9/0.3 (L eye), 0.7/0.2 (R eye), 0.4/0.4 (L lens), 0.3/0.3 (R lens), 1.0/0.8 (L Optical Nerve), 0.5/0.3 (R Optical Nerve), 0.3/0.2 (L Cochlea), 0.1/0.1 (R Cochlea), 4.6/0.2 (brain), 2.4/0.1 (brain-PTV), 5.1/0.9 (PTV). Compared to Eclipse, which generated an average of 607 segments, the stratified plan reduced (−) or increased (+) the maximum/mean dose, in Gy, by −10.2/−4.1 (brainstem), −10.5/−8.9 (chiasm), +0.0/−0.1 (spinal cord), −4.9/−3.4 (L eye), −4.1/−2.5 (R eye), −2.8/−2.7 (L lens), −2.1/−1.9 (R lens), −7.6/−6.5 (L Optical Nerve), −8.9/−6.1 (R Optical Nerve), −1.3/−1.9 (L Cochlea), −1.8/−1.8 (R Cochlea), +1.7/−2.1 (brain), +3.2/−2.6 (brain-PTV), +1.8/+0.3 Gy (PTV. The stratified plan was also more homogeneous in the PTV. Conclusion: This novel solver can transform complicated fluence maps into significantly fewer deliverable MLC segments than the commercial system while achieving superior dosimetry. Funding support partially contributed by Varian.

60 APPLIED LIFE SCIENCES↗

TU-F-17A-04: Respiratory Phase-Resolved 3D MRI with Isotropic High Spatial Resolution: Determination of the Average Breathing Motion Pattern for Abdominal Radiotherapy Planning

Purpose: To develop a retrospective 4D-MRI technique (respiratory phase-resolved 3D-MRI) for providing an accurate assessment of tumor motion secondary to respiration. Methods: A 3D projection reconstruction (PR) sequence with self-gating (SG) was developed for 4D-MRI on a 3.0T MRI scanner. The respiration-induced shift of the imaging target was recorded by SG signals acquired in the superior-inferior direction every 15 radial projections (i.e. temporal resolution 98 ms). A total of 73000 radial projections obtained in 8-min were retrospectively sorted into 10 time-domain evenly distributed respiratory phases based on the SG information. Ten 3D image sets were then reconstructed offline. The technique was validated on a motion phantom (gadolinium-doped water-filled box, frequency of 10 and 18 cycles/min) and humans (4 healthy and 2 patients with liver tumors). Imaging protocol included 8-min 4D-MRI followed by 1-min 2D-realtime (498 ms/frame) MRI as a reference. Results: The multiphase 3D image sets with isotropic high spatial resolution (1.56 mm) permits flexible image reformatting and visualization. No intra-phase motion-induced blurring was observed. Comparing to 2D-realtime, 4D-MRI yielded similar motion range (phantom: 10.46 vs. 11.27 mm; healthy subject: 25.20 vs. 17.9 mm; patient: 11.38 vs. 9.30 mm), reasonable displacement difference averaged over the 10 phases (0.74mm; 3.63mm; 1.65mm), and excellent cross-correlation (0.98; 0.96; 0.94) between the two displacement series. Conclusion: Our preliminary study has demonstrated that the 4D-MRI technique can provide high-quality respiratory phase-resolved 3D images that feature: a) isotropic high spatial resolution, b) a fixed scan time of 8 minutes, c) an accurate estimate of average motion pattern, and d) minimal intra-phase motion artifact. This approach has the potential to become a viable alternative solution to assess the impact of breathing on tumor motion and determine appropriate treatment margins. Comparison with 4D-CT in a clinical setting is warranted to assess the value of 4D-MRI in radiotherapy planning. This work supported in part by grant 1R03CA173273-01.

60 APPLIED LIFE SCIENCES↗

Using computer-extracted image features for modeling of error-making patterns in detection of mammographic masses among radiology residents

Purpose: Mammography is the most widely accepted and utilized screening modality for early breast cancer detection. Providing high quality mammography education to radiology trainees is essential, since excellent interpretation skills are needed to ensure the highest benefit of screening mammography for patients. The authors have previously proposed a computer-aided education system based on trainee models. Those models relate human-assessed image characteristics to trainee error. In this study, the authors propose to build trainee models that utilize features automatically extracted from images using computer vision algorithms to predict likelihood of missing each mass by the trainee. This computer vision-based approach to trainee modeling will allow for automatically searching large databases of mammograms in order to identify challenging cases for each trainee. Methods: The authors’ algorithm for predicting the likelihood of missing a mass consists of three steps. First, a mammogram is segmented into air, pectoral muscle, fatty tissue, dense tissue, and mass using automated segmentation algorithms. Second, 43 features are extracted using computer vision algorithms for each abnormality identified by experts. Third, error-making models (classifiers) are applied to predict the likelihood of trainees missing the abnormality based on the extracted features. The models are developed individually for each trainee using his/her previous reading data. The authors evaluated the predictive performance of the proposed algorithm using data from a reader study in which 10 subjects (7 residents and 3 novices) and 3 experts read 100 mammographic cases. Receiver operating characteristic (ROC) methodology was applied for the evaluation. Results: The average area under the ROC curve (AUC) of the error-making models for the task of predicting which masses will be detected and which will be missed was 0.607 (95% CI,0.564-0.650). This value was statistically significantly different from 0.5 (p < 0.0001). For the 7 residents only, the AUC performance of the models was 0.590 (95% CI,0.537-0.642) and was also significantly higher than 0.5 (p = 0.0009). Therefore, generally the authors’ models were able to predict which masses were detected and which were missed better than chance. Conclusions: The authors proposed an algorithm that was able to predict which masses will be detected and which will be missed by each individual trainee. This confirms existence of error-making patterns in the detection of masses among radiology trainees. Furthermore, the proposed methodology will allow for the optimized selection of difficult cases for the trainees in an automatic and efficient manner.

60 APPLIED LIFE SCIENCES↗

Dynamically patterning x-ray beam by a femtosecond optical laser

Modern science and technology have greatly benefitted from our ability to precisely manipulate light waves, in both their spatial and temporal degrees of freedom. In the x-ray region, however, spatial control has been virtually static mainly due to stringent requirements for realizing high-performance optical elements. The lack of dynamic spatial control of x-ray beam has prevented researchers from realizing more sophisticated use of the wave field, which has rapidly advanced in the optical region in the past decades. In this study, we propose a practical scheme to dynamically control local x-ray reflectivity of a perfect silicon crystal by a femtosecond optical laser and demonstrate a programmable spatial x-ray modulator. Our modulator aims for spatial manipulation of the x-ray amplitude and is shown to produce arbitrary grayscale patterns with spatial frequencies up to 25 per millimeter. The proposed modulation scheme opens up a platform to enable advanced x-ray sensing and imaging techniques that can fully harness the wave nature of x-rays.

47 OTHER INSTRUMENTATION↗

Human activities shape global patterns of decomposition rates in rivers

Rivers and streams contribute to global carbon cycling by decomposing immense quantities of terrestrial plant matter. However, decomposition rates are highly variable and large-scale patterns and drivers of this process remain poorly understood. Using a cellulose-based assay to reflect the primary constituent of plant detritus, we generated a predictive model (81% variance explained) for cellulose decomposition rates across 514 globally distributed streams. A large number of variables were important for predicting decomposition, highlighting the complexity of this process at the global scale. Predicted cellulose decomposition rates, when combined with genus-level litter quality attributes, explain published leaf litter decomposition rates with high accuracy (70% variance explained). Finally, our global map provides estimates of rates across vast understudied areas of Earth and reveals rapid decomposition across continental-scale areas dominated by human activities.

54 ENVIRONMENTAL SCIENCES↗

A DNA molecular printer capable of programmable positioning and patterning in two dimensions

Nanoscale manipulation and patterning usually require costly and sensitive top-down techniques such as those used in scanning probe microscopies or in semiconductor lithography. DNA nanotechnology enables exploration of bottom-up fabrication and has previously been used to design self-assembling components capable of linear and rotary motion. In this work, we combine three independently controllable DNA origami linear actuators to create a nanoscale robotic printer. The two-axis positioning mechanism comprises a moveable gantry, running on parallel rails, threading a mobile sleeve. We show that the device is capable of reversibly positioning a write head over a canvas through the addition of signaling oligonucleotides. We demonstrate “write” functionality by using the head to catalyze a local DNA strand–exchange reaction, selectively modifying pixels on a canvas. Furthermore, this work demonstrates the power of DNA nanotechnology for creating nanoscale robotic components and could find application in surface manufacturing, biophysical studies, and templated chemistry.

77 NANOSCIENCE AND NANOTECHNOLOGY↗

A general model for retroviral capsid pattern recognition by TRIM5 proteins

Restriction factors are intrinsic cellular defense proteins that have evolved to block microbial infections. Retroviruses such as HIV-1 are restricted by TRIM5 proteins, which recognize the viral capsid shell that surrounds, organizes, and protects the viral genome. TRIM5α uses a SPRY domain to bind capsids with low intrinsic affinity ( K D >1 mM), and therefore requires higher-order assembly into a hexagonal lattice to generate sufficient avidity for productive capsid recognition. TRIMCyp, on the other hand, binds HIV-1 capsids through a cyclophilin A domain, which has a well-defined binding site and higher ( K D ~10 μM) affinity for isolated capsid subunits. It has therefore been argued that TRIMCyp proteins may have dispensed with the need for higher-order assembly to function as antiviral factors. Here, we show that, consistent with its high degree of sequence similarity with TRIM5α, the TRIMCyp B-box 2 domain shares the same ability to self-associate and facilitate assembly of a TRIMCyp hexagonal lattice that can wrap about the HIV-1 capsid. We also show that under stringent experimental conditions, TRIMCyp-mediated restriction of HIV-1 is indeed dependent on higher-order assembly. Both forms of TRIM5 therefore use the same mechanism of avidity-driven capsid pattern recognition. IMPORTANCE Rhesus macaques and owl monkeys are highly resistant to HIV-1 infection due to the activity of TRIM5 restriction factors. The rhesus macaque TRIM5α protein blocks HIV-1 through a mechanism that requires self-assembly of a hexagonal TRIM5α lattice around the invading viral core. Lattice assembly amplifies very weak interactions between the TRIM5α SPRY domain and the HIV-1 capsid. Assembly also promotes dimerization of the TRIM5α RING E3 ligase domain, resulting in synthesis of polyubiquitin chains that mediate downstream steps of restriction. In contrast to rhesus TRIM5α, the owl monkey TRIM5 homolog, TRIMCyp, binds isolated HIV-1 CA subunits more tightly through its cyclophilin A domain, and was therefore thought to act independent of higher-order assembly. Here, we show that TRIMCyp shares the assembly properties of TRIM5α and that both forms of TRIM5 use the same mechanism of hexagonal lattice formation to promote viral recognition and restriction.

Wagner, Jonathan M.↗

Patterns in wetland microbial community composition and functional gene repertoire associated with methane emissions

Wetland restoration on peat islands previously drained for agriculture has potential to reverse land subsidence and sequester atmospheric carbon dioxide as peat accretes. However, the emission of methane could potentially offset the greenhouse gas benefits of captured carbon. As microbial communities play a key role in governing wetland greenhouse gas fluxes, we are interested in how microbial community composition and functions are associated with wetland hydrology, biogeochemistry, and methane emission, which is critical to modeling the microbial component in wetland methane fluxes and to managing restoration projects for maximal carbon sequestration. Here, we couple sequence-based methods with biogeochemical and greenhouse gas measurements to interrogate microbial communities from a pilot-scale restored wetland in the Sacramento-San Joaquin Delta of California, revealing considerable spatial heterogeneity even within this relatively small site. A number of microbial populations and functions showed strong correlations with electron acceptor availability and methane production; some also showed a preference for association with plant roots. Marker gene phylogenies revealed a diversity of major methane-producing and -consuming populations and suggested novel diversity within methanotrophs. Methanogenic archaea were observed in all samples, as were nitrate-, sulfate-, and metal-reducing bacteria, indicating that no single terminal electron acceptor was preferred despite differences in energetic favorability and suggesting spatial microheterogeneity and microniches. Notably, methanogens were negatively correlated with nitrate-, sulfate-, and metal-reducing bacteria and were most abundant at sampling sites with high peat accretion and low electron acceptor availability, where methane production was highest. Wetlands are the largest nonanthropogenic source of atmospheric methane but also a key global carbon reservoir. Characterizing belowground microbial communities that mediate carbon cycling in wetlands is critical to accurately predicting their responses to changes in land management and climate. Here, we studied a restored wetland and revealed substantial spatial heterogeneity in biogeochemistry, methane production, and microbial communities, largely associated with the wetland hydraulic design. We observed patterns in microbial community composition and functions correlated with biogeochemistry and methane production, including diverse microorganisms involved in methane production and consumption. We found that methanogenesis gene abundance is inversely correlated with genes from pathways exploiting other electron acceptors, yet the ubiquitous presence of genes from all these pathways suggests that diverse electron acceptors contribute to the energetic balance of the ecosystem. These investigations represent an important step toward effective management of wetlands to reduce methane flux to the atmosphere and enhance belowground carbon storage.

54 ENVIRONMENTAL SCIENCES↗

Fungal-Bacterial Cooccurrence Patterns Differ between Arbuscular Mycorrhizal Fungi and Nonmycorrhizal Fungi across Soil Niches

Soil bacteria and fungi are known to form niche-specific communities that differ between actively growing and decaying roots. Yet almost nothing is known about the cross-kingdom interactions that frame these communities and the environmental filtering that defines these potentially friendly or competing neighbors. We explored the temporal and spatial patterns of soil fungal (mycorrhizal and nonmycorrhizal) and bacterial cooccurrence near roots of wild oat grass, Avena fatua, growing in its naturalized soil in a greenhouse experiment. Amplicon sequences of the fungal internal transcribed spacer (ITS) and bacterial 16S rRNA genes from rhizosphere and bulk soils collected at multiple plant growth stages were used to construct covariation-based networks as a step toward identifying fungal-bacterial associations. Corresponding stable-isotope-enabled metagenome-assembled genomes (MAGs) of bacteria identified in cooccurrence networks were used to inform potential mechanisms underlying the observed links. Bacterial-fungal networks were significantly different in rhizosphere versus bulk soils and between arbuscular mycorrhizal fungi (AMF) and nonmycorrhizal fungi. Over 12 weeks of plant growth, nonmycorrhizal fungi formed increasingly complex networks with bacteria in rhizosphere soils, while AMF more frequently formed networks with bacteria in bulk soils. Analysis of network-associated bacterial MAGs suggests that some of the fungal-bacterial links that we identified are potential indicators of bacterial breakdown and consumption of fungal biomass, while others intimate shared ecological niches.

54 ENVIRONMENTAL SCIENCES↗

16S rRNA/rRNA Gene Ratios and Cell Activity Staining Reveal Consistent Patterns of Microbial Activity in Plant-Associated Soil

Although the majority of microorganisms in natural ecosystems are dormant, relatively little is known about the dynamics of the active and dormant microbial pools through both space and time. The limited knowledge of microbial activity-dormancy dynamics is in part due to uncertainty in the methods currently used to quantify active taxa. Here, we directly compared two of the most common methods (16S ratios and active cell staining) for estimating microbial activity in plant-associated soil and found that they were largely in agreement in the overarching patterns. Our results suggest that 16S ratios and active cell staining provide complementary information for measuring and interpreting microbial activity-dormancy dynamics in soils. They also support the idea that 16S rRNA/rRNA gene ratios have comparative value and offer a high-throughput, sequencing-based option for understanding relative changes in microbiome activity, as long as this method is coupled with quantification of community size.

16S rRNA↗

Proteomic-based stable isotope probing reveals taxonomically Distinct Patterns in Amino Acid Assimilation by Coastal Marine Bacterioplankton

Heterotrophic marine bacterioplankton are a critical component of the carbon cycle, processing nearly a quarter of annual global primary production, yet defining how substrate utilization preferences and resource partitioning structure these microbial communities remains a challenge. In this study, we utilized proteomics-based stable isotope probing (proteomic SIP) to characterize the assimilation of amino acids by coastal marine bacterioplankton populations. We incubated microcosms of seawater collected from Newport, OR and Monterey Bay, CA with 1 M 13 C-amino acids for 15 and 32 hours. Subsequent analysis of 13 C incorporation into protein biomass quantified the frequency and extent of isotope enrichment for identified proteins. Using these metrics we tested whether amino acid assimilation patterns were different for specific bacterioplankton populations. Proteins associated with Rhodobacterales and Alteromonadales tended to have a significantly high number of tandem mass spectra from 13 C-enriched peptides, while Flavobacteriales and SAR11 proteins generally had significantly low numbers of 13 C-enriched spectra. Rhodobacterales proteins associated with amino acid transport and metabolism had an increased frequency of 13 C-enriched spectra at time-point 2, while Alteromonadales ribosomal proteins were 13 C- enriched across time-points. Overall, proteomic SIP facilitated quantitative comparisons of dissolved free amino acids assimilation by specific taxa, both between sympatric populations and between protein functional groups within discrete populations, allowing an unprecedented examination of population-level metabolic responses to resource acquisition in complex microbial communities.

60 APPLIED LIFE SCIENCES↗