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At least 1,603 records · Page 89

Long-term compost amendment modulates wheat genotype differences in belowground carbon allocation, microbial rhizosphere recruitment and nitrogen acquisition

The implementation of soil health-promoting practices, such as cover cropping and compost application, has important implications for nutrient cycling and management in agroecosystems. At the same time, plant belowground carbon (C) allocation patterns can influence nutrient cycling and availability in soil through changes to the microbial community, but the effects may depend on the crop genotype and management practices in place. We evaluated belowground C allocation patterns using 13 C labeling and root architecture in two genotypes of winter wheat (Triticum aestivum) with different levels of exudation and belowground allocation strategies in soils with contrasting compost amendment legacy (108.7 Mg ha -1 every 2 years over 10 years vs. no compost). We also measured microbial community structure and function in the rhizosphere and quantified uptake of residue-derived N from 15 N-labelled cover crop residues. We found an interactive effect between soil management and genotype, where in the no-compost soil, the high-exudation genotype (Snowmass) increased exudation by over 4-fold, while the low-exudate genotype (Byrd) increased only 2-fold. While we did not observe genotype differences in rhizosphere enzyme activity or dissolved N pools, residue N uptake was 1.8 times greater for Snowmass in the compost-amended soil. There were more rhizosphere microbial taxa associated with the high-exudate genotype (Snowmass); nine bacterial and seven fungal families were indicative of Snowmass, versus one bacterial and four fungal families for Byrd. Our results suggest that the high-exudation strategy can influence the rhizosphere microbial community, and lead to greater short-term residue N uptake in high SOM soil. By directly linking root architecture, exudation, microbial communities, and N mineralization and uptake dynamics, this work demonstrates that plasticity in root C allocation is genotype-specific and influences microbial communities and nutrient cycling depending on the soil health context.

59 BASIC BIOLOGICAL SCIENCES↗

Warming, permafrost thaw and increased nitrogen availability as drivers for plant composition and growth across the Tibetan Plateau

Permafrost-affected ecosystems are subject to warming and thawing, which can increase the availability of subsurface nitrogen (N) with consequences in otherwise N-limited tundra and alpine vegetation. Here, we quantify the extent of warming and permafrost thawing and the corresponding effects on nitrogen availability and plant growth based on a 20-year survey across 14 sites on the Tibetan Plateau. The survey showed that most sites have been subject to warming and thawing and that the upper permafrost zone across all sites was rich in inorganic N, mainly as ammonium. We further explore the efficiency of plants to utilize 15 N-labelled inorganic N over five years following 15 N addition at the permafrost table far below the main root zone. The 15 N experiment showed that deep-rooted plant species were able to utilize the labelled N. A SEM model suggests that changes in vegetation can be explained by both active layer warming and permafrost thawing and the associated changes in inorganic nitrogen availability. Our results highlight a feedback mechanism of climate warming, in which released plant-available N may favour deep-rooted plants. This can explain important changes in plant composition and growth across the sites on the Tibetan Plateau.

54 ENVIRONMENTAL SCIENCES↗

Arabidopsis ABCG14 forms a homodimeric transporter for multiple cytokinins and mediates long-distance transport of isopentenyladenine-type cytokinins

Phytohormone cytokinins (CKs), primarily including trans-zeatin (tZ) and isopentenyladenine (iP) types, play critical roles in plant growth, development, and stress response. Long-distance transport of trans-zeatin (tZ)-type CKs has been well elucidated. However, long-distance transport of isopentenyladenine (iP)-type CKs remains unknown. Here, we demonstrated that the physiologically characterized ABC transporter AtABCG14 is also involved in long-distance transport of iP-type CKs. Biochemical characterization of AtABCG14 revealed that it forms homodimers and exhibits transporter properties towards multiple CKs, including iP and tZ types. AtABCG14 expression was induced by iP-type CKs in wild type and cyp735a1/a2 (cypDM) double mutant. The triple mutant atabcg14 cypDM exhibited more severe developmental phenotypes than atabcg14 and cypDM. CK quantification and 2 H 6 -iP isotope tracer experiment showed that the root-to-shoot transport of iP-type CKs was suppressed in atabcg14 cypDM. The reciprocal grafting of atabcg14 cypDM and cypDM revealed that AtABCG14 is responsible for the root xylem-loading and shoot phloem-distribution in the shootward transport of iP-type cytokinoins. Additionally, 2 H 6 -iP tracer experiment showed that the rootward transport of iP-type CKs was dependent on AtABCG14 expression in the roots. Collectively, our results demonstrate the long-distance circular transport of iP-type CKs mediated by AtABCG14 and provide new insights into the transport of CKs and other phytohormones.

59 BASIC BIOLOGICAL SCIENCES↗

Temporal variabilities of soil carbon dioxide fluxes from cornfield impacted by temperature and precipitation changes through high-frequent measurement and DAYCENT modelling

Soil carbon dioxide (CO 2 ) emissions from the field of corn (Zea mays L.) play an important role in global warming. This study investigated temporal variability of soil CO 2 fluxes (R s ) with soil temperature (T s ) and moisture (θ) and built DAYCENT models for predicting future impacts of climate changes on R s using the measured high-frequency data. R s trend was tested by Mann–Kendall and Sen Estimator. Predicted R s s under different climate scenarios were compared using Parallel-line Analysis. Herein, the findings indicated that daily R s exponentially increased with T s constrained by θ. During the θ of 27–31%, there was a strong exponential relationship between R s and T s , but the relationship was weaker for the θ of 38–41% and 22–26%. Soil environmental index (SEI, T s × θ) significantly impacted R s with linear regression R s 0.5 = 0.4599 + 0.002059 × SEI in 2008, 2009 and 2011. At the diurnal scale, there were different trends in R s s and relationships among R s and T s and θ in different years. Predicted yearly R s s, root R s s and corn yield in 2014–2049 increased with an increase in temperature scenarios, but the R s s significantly increased as temperature rose by 1°C or higher. Predicted yearly R s s, root R s s and yield reduced with precipitation scenario increase, and the root R s s and yield significantly diminished as precipitation increased by 15 and 30%. Predicted yearly R s from cornfields had a significantly increasing trend. Future research is needed to explore methods for mitigating cornfield R s and evaluating sensitivities of different cropland R s s to temperature changes.

54 ENVIRONMENTAL SCIENCES↗

Enhanced Spatial Proteomics and Metabolomics from a Single Tissue Section Using MALDI-MSI and LCM-microPOTS Platforms

Spatially resolved mass spectrometry (MS)-based multi-omics workflows are becoming more utilized for revealing the complex biology that occurs within tissues. However, these approaches commonly require multiple independent tissue sections to analyze the metabolite and protein compositions of these samples. This poses a significant challenge in preserving cell- or region-specific molecular fidelity, as variations between tissue sections can compromise the accurate correlation of molecular data. Here, in this study, we developed workflows for comprehensive multi-omics profiling from a single tissue section (STS) using different MS modalities. We enhanced the functionality of an electrically insulated substrate by employing metal-assisted approaches that enabled both MS-based untargeted spatial metabolomics and proteomics from STS. This allowed metabolite imaging using matrix-assisted laser desorption/ionization-MS imaging (MALDI-MSI), without compromising it for subsequent proteome profiling with laser capture microdissection (LCM)-based technology. Specifically, implementing copper tape as a backing for polyethylene naphthalate (PEN) slides enabled the detection of >140 metabolites across a poplar root tissue section using MALDI-trapped ion mobility spectrometry time of flight (timsTOF)-MS. Afterwards, we detected 6,571 unique proteins from two distinct root regions by leveraging LCM technology coupled to our microdroplet based sample preparation approach. We also developed an alternative workflow utilizing gold-coated PEN substrates for imaging with MALDI-Fourier-transform ion cyclotron resonance (FTICR)-MS, which permitted the profiling of >170 metabolites and the identification of 6,542 unique proteins across a single poplar root tissue section. These results were comparable to using each assay independently without modifications. These approaches offer new opportunities for high-resolution molecular profiling of multiple omics-levels across biological tissues.

Veličković, Marija [Pacific Northwest National Lab↗

Tracking Excited States in Wave Function Optimization Using Density Matrices and Variational Principles

We introduce an approach for finding individual excited states' energy stationary points in complete active space self-consistent field theory that is compatible with standard optimization methods and highly effective at overcoming difficulties due to root flipping and near-degeneracies. Inspired by both the maximum overlap method and recent progress in excited-state variational principles, our approach combines these ideas in order to track individual excited states throughout the orbital optimization process. In a series of tests involving root flipping, near-degeneracies, charge transfers, and double excitations, we show that this approach is more effective for state-specific optimization than either the naive selection of roots on the basis of energy ordering or a more direct generalization of the maximum overlap method. We offer evidence that this state-specific approach improves the performance of complete active space perturbation theory for vertical excitation energies. Furthermore, we find that the state-specific optimization can help avoid state-averaging-induced discontinuities on potential energy surfaces. With a simple implementation, a low cost, and compatibility with large active space methods, the approach is designed to be useful in a wide range of excited-state investigations.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Potassium and Calcium Isotopic Fractionation by Plants (Soybean [Glycine max], Rice [Oryza sativa], and Wheat [Triticum aestivum])

Here, we conducted hydroponic experiments growing soybean (Glycine max), rice (Oryza sativa), and wheat (Triticum aestivum) under K and Ca replete conditions to establish the degree of K isotopic fractionation by plants, and compare the isotopic fractionation of Ca and K. Each of the test plants displays fractionation relative to the growth solution favoring the light isotopes of K and Ca. The average δ 41 K values of the roots from the three plant species were similar, and have an overall average of -0.55 ± 0.24‰ 2s, while the overall average δ 44 Ca for roots is -0.67 ± 0.44. For leaves, the overall average of δ 41 K is -0.97 ± 0.4‰, compared to an overall average leaf δ 44 Ca of -0.83 ± 0.09‰. In the case of the soybean plants, the lightest K and Ca occurs in the stems with average δ 41 K of -1.31 ± 0.40‰ 2s and average δ 44 Ca of -1.20 ± 0.19 ‰ 2s. We present a simple box model involving the relative fluxes of K and its isotopic fractionation that reproduces our K isotopic observations and suggests a fractionation of ~0.8‰ with K uptake from solution by roots. Finally, directly comparing the per amu fractionation of K and Ca reveals an average factor of 2.05 ± 0.50 2s greater fractionation of K isotopes which may reflect their different roles and behaviors in plants.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Using Stable Carbon Isotopes of Seasonal Ecosystem Respiration to Determine Permafrost Carbon Loss

High latitude warming and permafrost thaw will expose vast stores of deep soil organic carbon (SOC) to decomposition. Thaw also changes water movement causing either wetter or drier soil. The fate of deep SOC under different thaw and moisture conditions is unclear. We measured weekly growing-season δ 13 C of ecosystem respiration (Recoδ 13 C) across thaw and moisture conditions (Shallow-Dry; Deep-Dry; Deep-Wet) in a soil warming manipulation. Deep SOC loss was inferred from known δ 13 C signatures of plant shoot, root, surface soil, and deep soil respiration. In addition, a two-year-old vegetation removal treatment (No Veg) was used to isolate surface and deep SOC decomposition contributions to Reco. In No Veg, seasonal Recoδ 13 C indicated that deep SOC loss increased as the soil column thawed while in vegetated areas root contributions appeared to dominate Reco. The Recoδ 13 C differences between Shallow-Dry and Deep-Dry were significant but surprisingly small. This most likely suggests that, under dry conditions, soil-warming stimulates root and surface SOC respiration with a negative 13 C signature that opposes the more positive 13 C signal from increased deep SOC respiration. In Deep-Wet conditions Recoδ 13 C suggests reduced deep SOC loss but could also reflect altered diffusion or methane (CH 4 ) dynamics. Together these results demonstrate that frequent Recoδ 13 C measurements can detect deep SOC loss, and that plants confound the signal. In conclusion, soil profile δ 13 C measurements, vegetation removal across thaw gradients, and isotopic effects of CH 4 dynamics could further deconvolute deep SOC loss via surface Reco.

54 ENVIRONMENTAL SCIENCES↗

Critical Zone Water Balance Over 13 Years in a Semiarid Savanna

Abstract Quantifying how much and when precipitation (P) becomes runoff (R), evapotranspiration (ET), and drainage from the root zone (D) is key to understanding how climate and land use impact hydrology of the critical zone. We quantify water balance dynamics of a semiarid savanna with a summer/winter rainfall pattern with 13 years of water fluxes and soil moisture. We find multiyear P is partitioned 96% to ET and 7% to R, while D (−3%) is negligible when considering measurement uncertainty. While weather regulates ET over diurnal time scales, soil water inputs control seasonal to annual ET amounts. Seasonal water availability, estimated by soil moisture inputs, is more closely tracked by ET rather than time‐averaged soil moisture or P. Surprisingly, we find significant, episodic carryover of soil moisture from the summer to spring growing season. Abundant late‐summer P can supply ET in the subsequent spring, even after multimonth dry periods. However, over an annual cycle beginning in early summer, nearly all soil moisture is used by ET. Likewise, D beyond the monitored root zone, assisted by downward hydraulic distribution in plant roots, occurs within a season, but this is counteracted by subsequent ET extraction of deep moisture over the year. Thus, negligible long‐term D occurs, though there is considerable uncertainty in estimation of this small flux as the residual of much larger ones. These comprehensive, long‐term measurements support expectations about the overriding importance of ET in the dryland critical zone water balance and reveal an unexpected degree of interseasonal water storage.

Scott, Russell L.↗

The importance of trait selection in ecology

Two recent publications have explored the importance of roots for understanding plant form and function but reached opposite conclusions on the basis of largely overlapping data. Carmona et al. concluded that their results “do not confirm the strong covariation between leaf and fine-root traits predicted by the plant economics spectrum hypothesis”. By contrast, Weigelt et al. concluded that “key leaf and fine-root traits were aligned along the expected [fast–slow] ‘conservation’ gradient of plant economic investment”. Furthermore, here we reflect on the causes for the apparent discrepancies of these studies and show that the rationale behind trait selection is vital for the conclusions.

59 BASIC BIOLOGICAL SCIENCES↗

The mutualism effector MiSSP7 of Laccaria bicolor alters the interactions between the poplar JAZ6 protein and its associated proteins

Despite the pivotal role of jasmonic acid in the outcome of plant-microorganism interactions, JA-signaling components in roots of perennial trees like western balsam poplar (Populus trichocarpa) are poorly characterized. Here we decipher the poplar-root JA-perception complex centered on PtJAZ6, a co-repressor of JA-signaling targeted by the effector protein MiSSP7 from the ectomycorrhizal basidiomycete Laccaria bicolor during symbiotic development. Through protein–protein interaction studies in yeast we determined the poplar root proteins interacting with PtJAZ6. Moreover, we assessed via yeast triple-hybrid how the mutualistic effector MiSSP7 reshapes the association between PtJAZ6 and its partner proteins. In the absence of the symbiotic effector, PtJAZ6 interacts with the transcription factors PtMYC2s and PtJAM1.1. In addition, PtJAZ6 interacts with it-self and with other Populus JAZ proteins. Finally, MiSSP7 strengthens the binding of PtJAZ6 to PtMYC2.1 and antagonizes PtJAZ6 homo-/heterodimerization. We conclude that a symbiotic effector secreted by a mutualistic fungus may promote the symbiotic interaction through altered dynamics of a JA-signaling-associated protein–protein interaction network, maintaining the repression of PtMYC2.1-regulated genes.

59 BASIC BIOLOGICAL SCIENCES↗

Anti-fungal bioactive terpenoids in the bioenergy crop switchgrass (Panicum virgatum) may contribute to ecotype-specific microbiome composition

Abstract Plant derived bioactive small molecules have attracted attention of scientists across fundamental and applied scientific disciplines. We seek to understand the influence of these phytochemicals on rhizosphere and root-associated fungi. We hypothesize that – consistent with accumulating evidence that switchgrass genotype impacts microbiome assembly – differential terpenoid accumulation contributes to switchgrass ecotype-specific microbiome composition. An initial in vitro Petri plate-based disc diffusion screen of 18 switchgrass root derived fungal isolates revealed differential responses to upland- and lowland-isolated metabolites. To identify specific fungal growth-modulating metabolites, we tested fractions from root extracts on three ecologically important fungal isolates – Linnemania elongata, Trichoderma sp. and Fusarium sp. Saponins and diterpenoids were identified as the most prominent antifungal metabolites. Finally, analysis of liquid chromatography-purified terpenoids revealed fungal inhibition structure – activity relationships (SAR). Saponin antifungal activity was primarily determined by the number of sugar moieties – saponins glycosylated at a single core position were inhibitory whereas saponins glycosylated at two core positions were inactive. Saponin core hydroxylation and acetylation were also associated with reduced activity. Diterpenoid activity required the presence of an intact furan ring for strong fungal growth inhibition. These results inform future breeding and biotechnology strategies for crop protection with reduced pesticide application.

09 BIOMASS FUELS↗

Large-scale replicated field study of maize rhizosphere identifies heritable microbes

Up until now, soil microbes that colonize plant roots and are responsive to differences in plant genotype remain to be ascertained for agronomically important crops. From a very large-scale longitudinal field study of 27 maize inbred lines planted in three fields, with partial replication 5 y later, we identify root-associated microbiota exhibiting reproducible associations with plant genotype. Analysis of 4,866 samples identified 143 operational taxonomic units (OTUs) whose variation in relative abundances across the samples was significantly regulated by plant genotype, and included five of seven core OTUs present in all samples. Plant genetic effects were significant amid the large effects of plant age on the rhizosphere microbiome, regardless of the specific community of each field, and despite microbiome responses to climate events. Seasonal patterns showed that the plant root microbiome is locally seeded, changes with plant growth, and responds to weather events. However, against this background of variation, specific taxa responded to differences in host genotype. If shown to have beneficial functions, microbes may be considered candidate traits for selective breeding.

59 BASIC BIOLOGICAL SCIENCES↗

Linked nitrogen and carbon dynamics reveal distinct pools and patterns in a deep, weathered bedrock rhizosphere

Nitrogen is one of the most limiting nutrients to forest productivity worldwide. Recently, it has been established that diverse ecosystems source a substantial fraction of their water from weathered bedrock, leading to questions about whether root-driven nitrogen cycling extends into weathered bedrock as well. In this study, we specifically examined nitrogen dynamics using specialized instrumentation distributed across a 16 m weathered bedrock vadose zone (WBVZ) underlying an old growth forest in northern California where the rhizosphere—composed of plant roots and their associated microbiome—extends meters into rock. We documented total dissolved nitrogen (TDN), dissolved organic carbon (DOC), inorganic nitrogen (ammonium and nitrate), and CO 2 and O 2 gases every 1.5 m to 16 m depth for 2 y. We found that TDN concentrations increased with depth, were an order of magnitude greater at 15 m than in the upper 30 cm, and that the majority of TDN throughout the weathered bedrock vadose zone was organic. We also found that TDN concentrations are influenced by depth, season, and interannual precipitation patterns. Carbon isotope composition of the DOC suggests that dissolved organic matter in the WBVZ is primarily derived from plant sources, and not the nitrogen-rich bedrock. We conclude that nitrogen dynamics in the WBVZ may be driven, in part, by an active rhizosphere, meters below the base of soil, and we argue that weathered bedrock horizons may play a key role in C-N cycling in ecosystems with deep-rooted plants.

Science & Technology - Other Topics↗

Comparison of three DWM-based wake models at above-rated wind speeds

In this study we investigate three mid-fidelity wind turbine wake models based on the dynamic wake meandering (DWM) model principle, and compare their performance with a reference dataset, produced with large-eddy simulations using the actuator line model. The models are compared with respect to flow field, power, and loads on a row of four 5MW reference turbines experiencing above-rated wind conditions. In general, the DWM models show fairly good agreement with large-eddy simulation for the time-averaged flow fields, blade forces and power, with increasing differences along the turbine row. Also when comparing fatigue loads of blade root moments, the differences between the models increase further into the row, with deviations up to 25 % of the reference case. However, while the development in blade root moment fatigue along the turbine row is predominantly driven by the energy content at the frequency corresponding to the turbine's rotational period (1 P ) for the DWM models, the large-eddy simulation results suggest that the key drivers for the blade root and tower loads are the increase in meandering and energy at higher frequencies (> 1 P ) deeper into the turbine row. For the tower loads, the DWM models highly underestimate the fatigue for the waked turbines. From these results, we suggest priorities for future model developments so that robust model implementations can be used in wind farm design and operation.

17 WIND ENERGY↗

Dynamic nitrogen fixation in an aerobic endophyte of Populus

Biological nitrogen fixation by microbial diazotrophs can contribute significantly to nitrogen availability in non-nodulating plant species. In this study of molecular mechanisms and gene expression relating to biological nitrogen fixation, the aerobic nitrogen-fixing endophyte Burkholderia vietnamiensis, strain WPB, isolated from Populus trichocarpa served as a model for endophyte–poplar interactions. Nitrogen-fixing activity was observed to be dynamic on nitrogen-free medium with a subset of colonies growing to form robust, raised globular like structures. Secondary ion mass spectrometry (NanoSIMS) confirmed that N-fixation was uneven within the population. A fluorescent transcriptional reporter (GFP) revealed that the nitrogenase subunit nifH is not uniformly expressed across genetically identical colonies of WPB and that only ~11% of the population was actively expressing the nifH gene. Higher nifH gene expression was observed in clustered cells through monitoring individual bacterial cells using single-molecule fluorescence in situ hybridization. Through 15 N 2 enrichment, we identified key nitrogenous metabolites and proteins synthesized by WPB and employed targeted metabolomics in active and inactive populations. We cocultivated WPB Pnif-GFP with poplar within a RhizoChip, a synthetic soil habitat, which enabled direct imaging of microbial nifH expression within root epidermal cells. We observed that nifH expression is localized to the root elongation zone where the strain forms a unique physical interaction with the root cells. This work employed comprehensive experimentation to identify novel mechanisms regulating both biological nitrogen fixation and beneficial plant–endophyte interactions.

15 N-tracking metabolomics↗

Spatiotemporal cytokinin response imaging and ISOPENTENYLTRANSFERASE 3 function in Medicago nodule development

Most legumes can establish a symbiotic association with soil rhizobia that trigger the development of root nodules. These nodules host the rhizobia and allow them to fix nitrogen efficiently. The perception of bacterial lipo-chitooligosaccharides (LCOs) in the epidermis initiates a signaling cascade that allows rhizobial intracellular infection in the root and de-differentiation and activation of cell division that gives rise to the nodule. Thus, nodule organogenesis and rhizobial infection need to be coupled in space and time for successful nodulation. The plant hormone cytokinin (CK) contributes to the coordination of this process, acting as an essential positive regulator of nodule organogenesis. However, the temporal regulation of tissue-specific CK signaling and biosynthesis in response to LCOs or Sinorhizobium meliloti inoculation in Medicago truncatula remains poorly understood. In this study, using a fluorescence-based CK sensor (pTCSn::nls:tGFP), we performed a high-resolution tissue-specific temporal characterization of the sequential activation of CK response during root infection and nodule development in M. truncatula after inoculation with S. meliloti. Loss-of-function mutants of the CK-biosynthetic gene ISOPENTENYLTRANSFERASE 3 (IPT3) showed impairment of nodulation, suggesting that IPT3 is required for nodule development in M. truncatula. Simultaneous live imaging of pIPT3::nls:tdTOMATO and the CK sensor showed that IPT3 induction in the pericycle at the base of nodule primordium contributes to CK biosynthesis, which in turn promotes expression of positive regulators of nodule organogenesis in M. truncatula.

59 BASIC BIOLOGICAL SCIENCES↗

It's only natural: Plant respiration in unmanaged systems

Abstract Respiration plays a key role in the terrestrial carbon cycle and is a fundamental metabolic process in all plant tissues and cells. We review respiration from the perspective of plants that grow in their natural habitat and how it is influenced by wide-ranging elements at different scales, from metabolic substrate availability to shifts in climate. Decades of field-based measurements have honed our understanding of the biological and environmental controls on leaf, root, stem, and whole-organism respiration. Despite this effort, there remain gaps in our knowledge within and across species and ecosystems, especially in more challenging-to-measure tissues like roots. Recent databases of respiration rates and associated leaf traits from species representing diverse biomes, plant functional types, and regional climates have allowed for a wider-lens view at modeling this important CO2 flux. We also re-analyze published data sets to show that maximum leaf respiration rates (R max) in species from around the globe are related both to leaf economic traits and environmental variables (precipitation and air temperature), but that root respiration does not follow the same latitudinal trends previously published for leaf data. We encourage the ecophysiological community to continue to expand their study of plant respiration in tissues that are difficult to measure and at the whole plant and ecosystem levels to address outstanding questions in the field.

59 BASIC BIOLOGICAL SCIENCES↗