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Mechanism and dynamics of fatty acid photodecarboxylase

Photoenzymes are rare biocatalysts driven by absorption of a photon at each catalytic cycle; they inspire development of artificial photoenzymes with valuable activities. Fatty acid photodecarboxylase (FAP) is a natural photoenzyme that has potential applications in the bio-based production of hydrocarbons, yet its mechanism is far from fully understood. RATIONALE To elucidate the mechanism of FAP, we studied the wild-type (WT) enzyme from Chlorella variabilis (CvFAP) and variants with altered active-site residues using a wealth of techniques, including static and time-resolved crystallography and spectroscopy, as well as biochemical and computational approaches. RESULTS A 1.8-Å-resolution CvFAP x-ray crystal structure revealed a dense hydrogen-bonding network positioning the fatty acid carboxyl group in the vicinity of the flavin adenine dinucleotide (FAD) cofactor. Structures solved from free electron laser and low-dose synchrotron x-ray crystal data further highlighted an unusual bent shape of the oxidized flavin chromophore, and showed that the bending angle (14°) did not change upon photon absorption (step 1) or throughout the photocycle. Calculations showed that bending substantially affected the energy levels of the flavin. Structural and spectroscopic analysis of WT and mutant proteins targeting two conserved active-site residues, R451 and C432, demonstrated that both residues were crucial for proper positioning of the substrate and water molecules and for oxidation of the fatty acid carboxylate by 1 FAD* (~300 ps in WT FAP) to form FAD ∙– (step 2). Time-resolved infrared spectroscopy demonstrated that decarboxylation occured quasi-instantaneously upon this forward electron transfer, consistent with barrierless bond cleavage predicted by quantum chemistry calculations and with snapshots obtained by time-resolved crystallography. Transient absorption spectroscopy in H 2 O and D 2 O buffers indicated that back electron transfer from FAD ∙– was coupled to and limited by transfer of an exchangeable proton or hydrogen atom (step 3). Unexpectedly, concomitant with FAD ∙– reoxidation (to a red-shifted form FAD RS ) in 100 ns, most of the CO 2 product was converted, most likely into bicarbonate (as inferred from FTIR spectra of the cryotrapped FAD RS intermediate). Calculations indicated that this catalytic transformation involved an active-site water molecule. Cryo-Fourier transform infrared spectroscopy studies suggested that bicarbonate formation (step 4) was preceded by deprotonation of an arginine residue (step 3). At room temperature, the remaining CO 2 left the protein in 1.5 μs (step 4'). The observation of residual electron density close to C432 in electron density maps derived from time-resolved and cryocrystallography data suggests that this residue may play a role in stabilizing CO 2 and/or bicarbonate. Three routes for alkane formation were identified by quantum chemistry calculations; the one shown in the figure is favored by the ensemble of experimental data. CONCLUSION Finally, we provide a detailed and comprehensive characterization of light-driven hydrocarbon formation by FAP, which uses a remarkably complex mechanism including unique catalytic steps. We anticipate that our results will help to expand the green chemistry toolkit.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

The ORNL Moderator Test Station Science Case

Oak Ridge National Laboratory (ORNL) hosts two world-leading slow neutron sources, the Spallation Neutron Source (SNS) and the High Flux Isotope Reactor (HFIR), and is currently developing the technical design for a Second Target Station (STS) for the SNS. Upon completion of the STS project, ORNL will be uniquely positioned to optimize each of its three neutron sources, the SNS First Target Station (FTS), the STS, and HFIR, in a complementary way. Among the essential aspects of a re-imagined FTS and the current STS design are high-brightness parahydrogen moderators—moderators which are optimized for high per-unit-area neutron brightness rather than integrated-across-large-area neutron intensity. The high-brightness moderators proposed for the STS will, for the brightness metric, significantly outperform the coupled moderators currently on the FTS for appropriately optimized neutron beamlines, provided the moderating hydrogen is converted to near-equilibrium levels of parahydrogen (approximately 99.8% at 20 K). The original FTS moderators, by contrast, were conservatively designed to be relatively insensitive to the exact ortho:para ratio, with a consequent loss in performance. As a result, a redesign of the FTS moderators assuming fully converted parahydrogen could result in significant performance improvements on the FTS coupled moderators, and more consistent performance over time for all hydrogen moderators. This “parahydrogen problem” is a long-standing challenge for the effective implementation of hydrogen cold moderators at high-power neutron sources. In addition, the development of new moderator concepts, whether based on previously unused materials, structured heterogeneous arrays, or even simply on changes in overall shape and size is significantly restricted at a large-scale production facility intended to use the resulting neutron beams. Accordingly, moderators for production neutron sources are often designed in a very conservative, low-risk fashion, even though this compromises the absolute neutronic performance. Advanced moderator concepts worthy of study include features that could not be tested without redesigning and redeploying the entire existing reflector, shielding, and neutron beamline installation, making such development efforts far more expensive than building a stand-alone test facility.

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗

Magnetocaloric refrigerator

The invention is for an apparatus and method for a refrigerator and a heat pump based on the magnetocaloric effect (MCE) offering a simpler, lighter, robust, more compact, environmentally compatible, and energy efficient alternative to traditional vapor-compression devices. The subject magnetocaloric apparatus alternately exposes a magnetocaloric material to strong and weak magnetic field while switching heat to and from the material. Action of the heat switches is coordinated with the magnetic field strength to move heat up the thermal gradient. The invention may be practiced with multiple magnetocaloric stages to attain large differences in temperature. Key applications include thermal management of electronics, as well as industrial and home refrigeration, heating, and air conditioning. The invention offers a simpler, lighter, compact, and robust apparatus compared to magnetocaloric devices of prior art. Furthermore, the invention may be run in reverse as a thermodynamic engine, receiving low-level heat and producing mechanical energy.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Regulation of bacterial stringent response by an evolutionarily conserved ribosomal protein L11 methylation

Lysine and arginine methylation is an important regulator of enzyme activity and transcription in eukaryotes. However, little is known about this covalent modification in bacteria. In this work, we investigated the role of methylation in bacteria. By reanalyzing a large phyloproteomics data set from 48 bacterial strains representing six phyla, we found that almost a quarter of the bacterial proteome is methylated. Many of these methylated proteins are conserved across diverse bacterial lineages, including those involved in central carbon metabolism and translation. Among the proteins with the most conserved methylation sites is ribosomal protein L11 (bL11). bL11 methylation has been a mystery for five decades, as the deletion of its methyltransferase PrmA causes no cell growth defects. Comparative proteomics analysis combined with inorganic polyphosphate and guanosine tetra/pentaphosphate assays of the ΔprmA mutant in Escherichia coli revealed that bL11 methylation is important for stringent response signaling. In the stationary phase, we found that the ΔprmA mutant has impaired guanosine tetra/pentaphosphate production. This leads to a reduction in inorganic polyphosphate levels, accumulation of RNA and ribosomal proteins, and an abnormal polysome profile. Overall, our investigation demonstrates that the evolutionarily conserved bL11 methylation is important for stringent response signaling and ribosomal activity regulation and turnover.

59 BASIC BIOLOGICAL SCIENCES↗

Addressing energy storage needs at lower cost via on-site thermal energy storage in buildings

Cost-effective energy storage is a critical enabler for the large-scale deployment of renewable electricity. Significant resources have been directed toward developing cost-effective energy storage, with research and development efforts dominated by work on lithium ion (Li-ion) battery technology. Though Li-ion batteries have many attractive qualities, it is not clear whether they can provide an affordable levelized cost of storage (LCOS) for certain applications, such as buildings. Buildings consume most of the world's electricity, and as much as 50% of their consumption goes toward meeting thermal loads. Thermal energy storage (TES) can provide a cost-effective alternative to Li-ion batteries for buildings; however, two questions remain to be answered. First, how much of total building energy storage requirements can be met via thermal storage for building loads? Second, can the LCOS for TES be favorable compared with Li-ion batteries? In this perspective, using the United States as a case study, we show that the total requirement for TES in buildings is in the range of ~1200–4500 electrical GW h, depending on the fraction of solar versus wind in the generation mix. Furthermore, we show that with at least 25% wind generation, all of the storage needed by buildings to support the grid can be met by TES. We also introduce a framework to calculate LCOS for on-site TES in buildings to enable a direct comparison with electrical storage technologies such as Li-ion batteries. This is not trivial, because the input energy type for TES (electricity) differs from the output energy type (thermal energy), and the efficiency can depend on ambient conditions. Our LCOS analysis shows that in many situations, TES can be more cost-effective for buildings than Li-ion batteries. We conclude our perspective by discussing future research and development opportunities that can significantly advance the deployment of TES for buildings to help enable a renewable electricity-dependent grid.

25 ENERGY STORAGE↗

EfgA is a conserved formaldehyde sensor that leads to bacterial growth arrest in response to elevated formaldehyde

Normal cellular processes give rise to toxic metabolites that cells must mitigate. Formaldehyde is a universal stressor and potent metabolic toxin that is generated in organisms from bacteria to humans. Methylotrophic bacteria such as Methylorubrum extorquens face an acute challenge due to their production of formaldehyde as an obligate central intermediate of single-carbon metabolism. Mechanisms to sense and respond to formaldehyde were speculated to exist in methylotrophs for decades but had never been discovered. Here, we identify a member of the DUF336 domain family, named efgA for enhanced formaldehyde growth, that plays an important role in endogenous formaldehyde stress response in M . extorquens PA1 and is found almost exclusively in methylotrophic taxa. Our experimental analyses reveal that EfgA is a formaldehyde sensor that rapidly arrests growth in response to elevated levels of formaldehyde. Heterologous expression of EfgA in Escherichia coli increases formaldehyde resistance, indicating that its interaction partners are widespread and conserved. EfgA represents the first example of a formaldehyde stress response system that does not involve enzymatic detoxification. Thus, EfgA comprises a unique stress response mechanism in bacteria, whereby a single protein directly senses elevated levels of a toxic intracellular metabolite and safeguards cells from potential damage.

59 BASIC BIOLOGICAL SCIENCES↗

Complete biosynthesis of salicylic acid from phenylalanine in plants

Salicylic acid (SA) is a pivotal phytohormone for plant responses to biotic and abiotic stresses. Plants have evolved two pathways to produce SA: the isochorismate synthase and phenylalanine ammonia lyase (PAL) pathways. Whereas the isochorismate synthase pathway has been fully identified, the PAL pathway remains incomplete. Here we report the full characterization of the PAL pathway for SA biosynthesis via functional analysis of rice (Oryza sativa) SA-DEFICIENT GENE 1 (OSD1) to OSD4. The cinnamoyl-coenzyme A (CoA) ligase OSD1 catalyses the conversion of trans-cinnamic acid to cinnamoyl-CoA, which is subsequently transformed to benzoyl-CoA via the β-oxidative pathway in peroxisomes. The resulting benzoyl-CoA is further converted to benzyl benzoate by the peroxisomal benzoyltransferase OSD2. Benzyl benzoate is subsequently hydroxylated to benzyl salicylate by the endoplasmic reticulum membrane-resident cytochrome P450 OSD3, which is ultimately hydrolysed to salicylic acid by the cytoplasmic carboxylesterase OSD4. Evolutionary analyses reveal that the PAL pathway was first assembled before the divergence of gymnosperms and has been conserved in most seed plants. Activation of the PAL pathway in rice significantly enhances salicylic acid levels and plant immunity. Completion of the PAL pathway provides critical insights into the primary salicylic acid biosynthetic pathway across plant species and offers a precise target for modulating crop immunity.

59 BASIC BIOLOGICAL SCIENCES↗

Four-dimensional phase space tomography from one-dimensional measurements of a hadron beam

In this paper, we use one-dimensional measurements to infer the four-dimensional phase space density of an accumulated proton beam in the Spallation Neutron Source (SNS) accelerator. The reconstruction was performed by maximizing the distribution’s entropy subject to the measurement constraints and thus represents the most conservative inference from the data. The reconstructed distribution reproduces the measured profiles down to the noise level, and simulations indicate that the problem is reasonably well constrained. Similar measurements could serve as benchmarks for beam dynamics simulations in the SNS or hadron accelerators.

43 PARTICLE ACCELERATORS↗

Energy Exascale Earth System Model v2.0

First release of version 2 of the Energy Exascale Earth System Model. The atmosphere component remains EAM. Major changes since version 1 include: all column-physics parameterizations are computed on a separate grid that has approximately half the number of points of the dynamics grid, a new nonhydrostatic dynamical core (running in hydrostatic mode) with semi-Lagrangian tracer transport, CLUBB updated from v1 to v2, a new convective trigger (dCAPE/ULL) based on the dynamic Convective Available Potential Energy (CAPE) (dCAPE) and the Unrestricted Launch Level (ULL) concepts is used in ZM. minimum cloud droplet number changed, gravity wave drag energy conservation fixed and new tunings used, dust emission size distribution changed to emit more coarse dust particles The land component is still ELM. Major changes since version 1 include: using SNICAR-AD for radiation in snow to match the sea-ice model and fixing bugs in snow compaction and water state calculation. The ocean component remains MPAS-ocean. Major change since version 1 include: Redi isopycnal mixing has been updated, tested, and tuned in combination with the Gent-McWilliams parameterization, a sign error was fixed in the 3rd-order flux routines, the mesh used in low-resolution coupled cases was modified and the time steps adjusted, new regionally refined meshes were created, one focused on North America and another on the Southern Ocean, including ice shelf cavities. The sea-ice component remains MPAS-seaice. Major changes since version 1 include: A new heat- and freshwater-conserving coupling of frazil ice, turning off of SSH filtering, addition of SNICAR-AD and snow grain aging. The land-ice component remains MPAS-Albany-landIce (MALI) and is a static ice sheet. There is more out-of-the-box support for cryosphere configurations including the new regionally refined configuration around Antarctica with ice shelf cavities. The river model is MOSART. The half-degree river mesh was redone so it no longer treats Black and Caspian seas as ocean. The coupler remains cpl7/MCT. Major changes since version 1 include handling of ice shelf melt fluxes (heat / freshwater exchange with the ocean), and data icebergs. All components allow regional refinement of their meshes and two separate example of refinement, one in and around North America and one around Antarctica and the Southern Ocean, are provided.

E3SM Project, DOE↗

Review of Pantex Radiation Safety Department 2020 Backup Dosimetry Processing Methodology

In the first quarter (Q1) of Calendar Year 2020, the Pantex Radiation Safety Department (Pantex) experienced various issues and failures with their dosimeter processing equipment. Efforts to repair the equipment resulted in limited functionality without the ability to process dosimeters assigned to personnel for monitoring dose. As a result of the impairments to the dosimeter processing equipment, Pantex called upon Nevada National Security Site (NNSS) for support, as their designated backup processor. In January of 2021, LLNL Subject Matter Experts in External Dosimetry started an NNSA-requested review of Pantex’s personnel doses in Q1 2020. Its focus was the technical validity of the means and methods used by Pantex and its conclusions and provide a level of confidence to National Nuclear Security Administration (NNSA) that the resulting personnel doses were reasonable, conservative, and defensible.

61 RADIATION PROTECTION AND DOSIMETRY↗

A Combined Water and CO 2 Direct Air Capture System (Final Technical Report)

The primary objective of the project was to demonstrate the technical and economic performance of a technology that simultaneously captures CO 2 and water from the air – a Hybrid Direct Air Capture system (HDAC). In HDAC, air is passed over water capture section as well as a CO 2 selective sorbent to remove CO 2 from the air stream. Combining potable water generation and CO 2 capture in a single device with the unique energy conserving features of the proposed design enables long-term projected CAPEX under $\$$750/t-CO 2 and levelized cost of capture (LCOC) of $\$$140/t-CO 2 . Project DE-FE-0031970 "A Combined Water and CO 2 Direct Air Capture System" ran for four years and three months from 10/01/2020 to 12/31/2024 for three budget periods and an extension period. The total project budget at completion was $\$$3,534,408 consisting of a $\$$2,680,064 federal share and $\$$854,344 cost share. The pilot plant was successfully designed, engineered, built and commissioned during the initial three budget periods. The plant demonstrated successful water capture during this period, whereas initial CO 2 capture was well below target values. The project team requested an extension period during which alternate materials and beds were evaluated. By the end of the project, both moisture swing adsorption of CO 2 and water capture had successfully been demonstrated at target capture rates making further scale-up of the technology viable.

42 ENGINEERING↗

Biophysical and Biochemical Characterization of TP0037, a D-Lactate Dehydrogenase, Supports an Acetogenic Energy Conservation Pathway in Treponema pallidum

A longstanding conundrum in Treponema pallidum biology concerns how the spirochete generates sufficient energy to fulfill its complex pathogenesis processes during human syphilitic infection. For decades, it has been assumed that the bacterium relies solely on glucose catabolism (via glycolysis) for generation of its ATP. However, the organism’s robust motility, believed to be essential for human tissue invasion and dissemination, would require abundant ATP likely not provided by the parsimony of glycolysis. As such, additional ATP generation, either via a chemiosmotic gradient, substrate-level phosphorylation, or both, likely exists in T. pallidum. Along these lines, we have hypothesized that T. pallidum exploits an acetogenic energy conservation pathway that relies on the redox chemistry of flavins. Central to this hypothesis is the apparent existence in T. pallidum of an acetogenic pathway for the conversion of d-lactate to acetate. Herein we have characterized the structural, biophysical, and biochemical properties of the first enzyme (d-lactate dehydrogenase [d-LDH]; TP0037) predicted in this pathway. Binding and enzymatic studies showed that recombinant TP0037 consumed d-lactate and NAD+ to produce pyruvate and NADH. The crystal structure of TP0037 revealed a fold similar to that of other d-acid dehydrogenases; residues in the cofactor-binding and active sites were homologous to those of other known d-LDHs. The crystal structure and solution biophysical experiments revealed the protein’s propensity to dimerize, akin to other d-LDHs. This study is the first to elucidate the enzymatic properties of T. pallidum’s d-LDH, thereby providing new compelling evidence for a flavin-dependent acetogenic energy conservation (ATP-generating) pathway in T. pallidum.

59 BASIC BIOLOGICAL SCIENCES↗

Anomalies in the topology of the temperature fluctuations in the cosmic microwave background: An analysis of the NPIPE and FFP10 data releases

We present a topological analysis of the temperature fluctuation maps from the Planck 2020 Data Release 4 NPIPE dataset and the Planck 2018 Data Release 3 FFP10 dataset. We performed a multiscale analysis in terms of the homology characteristics of the maps, invoking relative homology to account for the analysis in the presence of masks. We performed our analysis for a range of smoothing scales spanning sub- and super-horizon scales corresponding to a full width at half maximum (FWHM) of 5',10',20',40',80',160',320', and 640', and employed simulations based on the standard model for comparison, which assumes the initial fluctuation field to be an isotropic and homogeneous Gaussian random field. Examining the behavior of topological components, represented by the 0D homology group, we find the observations to be approximately 2σ or less deviant from the simulations for all resolutions and scales for the NPIPE dataset. For the FFP10 dataset, we detect a 2.96σ deviation between the observations and simulations at N = 128, FWHM = 80'. For the topological loops, represented by the first homology group, the simulations and observations are consistent within 2σ for most resolutions and scales for both the datasets. However, for the NPIPE dataset, we observe a high deviation between the observation and simulations in the number of loops at FWHM = 320', but at a low dimensionless threshold ν = –2.5. Under a Gaussian assumption, this would amount to a deviation of ~4σ. However, the distribution in this bin is manifestly non-Gaussian and does not obey Poisson statistics either. In the absence of a true theoretical understanding, we simply note that the significance is higher than what may be resolved by 600 simulations, yielding an empirical p-value of at most 0.0016. Specifically in this case, our tests indicate that the numbers arise from a statistically stable regime, despite being based on small numbers. For the FFP10 dataset, the differences are not as strong as for the NPIPE dataset, indicating a 2.77σ deviation at this resolution and threshold. The Euler characteristic, which is the alternating sum of the ranks of relative homology groups, reflects the deviations in the components and loops. To assess the significance of combined levels for a given scale, we employed the empirical and theoretical versions of the χ 2 test as well as the nonparametric Tukey depth test. Although all statistics exhibit a stable distribution, we favor the empirical version of the χ 2 test in the final interpretation, as it indicates the most conservative differences. For the NPIPE dataset, we find that the components and loops differ at more than 95%, but agree within the 99% confidence level with respect to the base model at N = 32, FWHM = 320'. The Euler characteristic at this resolution displays a per mil deviation. In contrast, the FFP10 dataset shows that the observations are consistent with the base model within the 95% confidence level, at this and smaller scales. This is consistent with the observations of the Planck analysis pipeline via Minkowski functionals. For the largest smoothing scale, N = 16, FWHM = 640', both datasets exhibit an anomalous behavior of the loops, where FFP10 data exhibit a deviation that is larger by an order of magnitude than that of the NPIPE dataset. In contrast, the values for the topological components and the Euler characteristic agree between observations and model to within a confidence level of 99%. However, for the largest scales, the statistics are based on low numbers and may have to be regarded with caution. Even though both datasets exhibit mild to significant discrepancies, they also exhibit contrasting behaviors at various instances. Therefore, we do not find it feasible to convincingly accept or reject the null hypothesis. Disregarding the large-scale anomalies that persist at similar scales in WMAP and Planck, observations of the cosmic microwave background are largely consistent with the standard cosmological model within 2σ.

79 ASTRONOMY AND ASTROPHYSICS↗

Rice Business Plan Competition (Final Report)

During the grant period, Rice University held a total of three regional Clean Energy Business Plan Competitions for graduate level students with a particular focus on attracting student startups in the clean energy space (clean tech, renewable energy, transportation, batteries, sustainability, etc.) Over the three years of the grant, the Rice Business Plan Competition awarded a total of three ${$}$50,000 DOE-sponsored prizes to the winning startups (${$}$150,000 total). The Rice Energy Business Plan Competition’s DOE Clean Tech competition leveraged the resources and infrastructure of the existing annual Rice Business Plan Competition (RBPC), which is the world’s largest and richest intercollegiate graduate-level business plan competition in the world

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Impacts of Model Building Energy Codes

The Department of Energy (DOE) Building Energy Codes Program (BECP) periodically evaluates state and national impacts associated with energy codes in residential and commercial buildings. Pacific Northwest National Laboratory (PNNL), funded by DOE, assessed the prospective impacts of national model building energy codes from 2010 through 2040. The model codes of interest in this report are the International Energy Conservation Code (IECC) for residential buildings and ASHRAE Standard 90.1 for commercial buildings. This report describes the methodology used for the assessment and presents the impacts in terms of energy savings, consumer cost savings, and reduced CO 2 emissions at the state and national levels.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Ecologically-Informed Precision Conservation: A framework for increasing biodiversity in intensively managed agricultural landscapes with minimal sacrifice in crop production

Conservation actions are urgently needed to tackle biodiversity loss in intensively managed agricultural landscapes. Production lands are usually heterogeneous and contain low-yield areas that can be set aside for biodiversity conservation without serious yield losses. Here, we introduce Ecologically-Informed Precision Conservation, a framework that integrates yield mapping and ecological theory to select the best areas to create new set-asides while ensuring high crop yields at the farm/landscape level. Long-term yield maps can be generated using globally available satellite data and basic information on field/farm crop yield from farmers. Ecological principles are then used to select the subset of areas with the highest potential for biodiversity conservation by prioritising those that increase connectivity, maximise habitat heterogeneity and decrease landscape grain size. Here, the created non-crop habitats can be permanent and thus ensure biodiversity support over time. In addition, agricultural management efficiency can be enhanced by improving field shapes. The framework provides the basis for a practical, user-friendly tool that informs all interested stakeholders on how to rationalise existing agricultural landscapes using already-existing farming systems and available technologies. High cost-effectiveness from an economic and conservation perspective, along with the creation of heterogeneous non-crop habitats, make our framework a promising solution to re-design agricultural landscapes.

Precision agriculture↗

Warming of alpine tundra enhances belowground production and shifts community towards resource acquisition traits

Abstract Climate warming is expected to stimulate plant growth in high‐elevation and high‐latitude ecosystems, significantly increasing aboveground net primary production (ANPP). However, the effects of simultaneous changes in temperature, snowmelt timing, and summer water availability on total net primary production (NPP)—and elucidation of both above‐ and belowground responses—remain an important area in need of further study. In particular, measures of belowground net primary productivity (BNPP) are required to understand whether ANPP changes reflect changes in allocation or are indicative of a whole plant NPP response. Further, plant functional traits provide a key way to scale from the individual plant to the community level and provide insight into drivers of NPP responses to environmental change. We used infrared heaters to warm an alpine plant community at Niwot Ridge, Colorado, and applied supplemental water to compensate for soil water loss induced by warming. We measured ANPP, BNPP, and leaf and root functional traits across treatments after 5 yr of continuous warming. Community‐level ANPP and total NPP (ANPP + BNPP) did not respond to heating or watering, but BNPP increased in response to heating. Heating decreased community‐level leaf dry matter content and increased total root length, indicating a shift in strategy from resource conservation to acquisition in response to warming. Water use efficiency (WUE) decreased with heating, suggesting alleviation of moisture constraints that may have enabled the plant community to increase productivity. Heating may have decreased WUE by melting snow earlier and creating more days early in the growing season with adequate soil moisture, but stimulated dry mass investment in roots as soils dried down later in the growing season. Overall, this study highlights how ANPP and BNPP responses to climate change can diverge, and encourages a closer examination of belowground processes, especially in alpine systems, where the majority of NPP occurs belowground.

54 ENVIRONMENTAL SCIENCES↗

Incorporate day-ahead robustness and real-time incentives for electricity market design

In this paper, we propose a two-stage electricity market framework to explore the participation of distributed energy resources (DERs) in a day-ahead (DA) market and a real-time (RT) market. The objective is to determine the optimal bidding strategies of the aggregated DERs in the DA market and generate online incentive signals for DER-owners to optimize the social-welfare taking into account network operational constraints. Distributionally robust optimization is used to explicitly incorporate data-based statistical information of renewable forecasts into the supply/demand decisions in the DA market. We evaluate the conservativeness of bidding strategies distinguished by different risk aversion settings. In the RT market, a bi-level time-varying optimization problem is proposed to design the online incentive signals to tradeoff the RT imbalance penalty for distribution system operators (DSOs) and the costs of individual DER-owners. This enables tracking their optimal dispatch to provide fast balancing services, in the presence of time-varying network states while satisfying the voltage regulation requirement. Simulation results on both DA wholesale market and RT balancing market demonstrate the necessity of this two-stage design, and its robustness to uncertainties, the performance of convergence, the tracking ability and the feasibility of the resulting network operations.

24 POWER TRANSMISSION AND DISTRIBUTION↗