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At least 109 records · Page 6

Optimization of Molecular Methods for Detecting Duckweed-Associated Bacteria

The bacterial colonization dynamics of plants can differ between phylogenetically similar bacterial strains and in the context of complex bacterial communities. Quantitative methods that can resolve closely related bacteria within complex communities can lead to a better understanding of plant–microbe interactions. However, current methods often lack the specificity to differentiate phylogenetically similar bacterial strains. In this study, we describe molecular strategies to study duckweed–associated bacteria. We first systematically optimized a bead-beating protocol to co-isolate nucleic acids simultaneously from duckweed and bacteria. We then developed a generic fingerprinting assay to detect bacteria present in duckweed samples. To detect specific duckweed–bacterium associations, we developed a genomics-based computational pipeline to generate bacterial strain-specific primers. These strain-specific primers differentiated bacterial strains from the same genus and enabled the detection of specific duckweed–bacterium associations present in a community context. Moreover, we used these strain-specific primers to quantify the bacterial colonization of duckweed by normalization to a plant reference gene and revealed differences in colonization levels between strains from the same genus. Lastly, confocal microscopy of inoculated duckweed further supported our PCR results and showed bacterial colonization of the duckweed root–frond interface and root interior. The molecular methods introduced in this work should enable the tracking and quantification of specific plant-microbe associations within plant-microbial communities.

59 BASIC BIOLOGICAL SCIENCES↗

Green-light supplementation for enhanced lettuce growth under red- and blue-light-emitting diodes

Plants will be an important component of future long-term space missions. Lighting systems for growing plants will need to be lightweight, reliable, and durable, and light-emitting diodes (LEDs) have these characteristics. Previous studies demonstrated that the combination of red and blue light was an effective light source for several crops. Yet the appearance of plants under red and blue lighting is purplish gray making visual assessment of any problems difficult. The addition of green light would make the plant leave appear green and normal similar to a natural setting under white light and may also offer a psychological benefit to the crew. Green supplemental lighting could also offer benefits, since green light can better penetrate the plant canopy and potentially increase plant growth by increasing photosynthesis from the leaves in the lower canopy. In this study, four light sources were tested: 1) red and blue LEDs (RB), 2) red and blue LEDs with green fluorescent lamps (RGB), 3) green fluorescent lamps (GF), and 4) cool-white fluorescent lamps (CWF), that provided 0%, 24%, 86%, and 51% of the total PPF in the green region of the spectrum, respectively. The addition of 24% green light (500 to 600 nm) to red and blue LEDs (RGB treatment) enhanced plant growth. The RGB treatment plants produced more biomass than the plants grown under the cool-white fluorescent lamps (CWF treatment), a commonly tested light source used as a broad-spectrum control.

Non-NASA Center↗

Plant physical defenses contribute to a latitudinal gradient in resistance to insect herbivory within a widespread perennial grass

Premise: Herbivore pressure can vary across the range of a species, resulting in different defensive strategies. If herbivory is greater at lower latitudes, plants may be better defended there, potentially driving a latitudinal gradient in defense. However, relationships that manifest across the entire range of a species may be confounded by differences within genetic subpopulations, which may obscure the drivers of these latitudinal gradients. Methods: We grew plants of the widespread perennial grass Panicum virgatum in a common garden that included genotypes from three genetic subpopulations spanning an 18.5° latitudinal gradient. We then assessed defensive strategies of these plants by measuring two physical resistance traits—leaf mass per area (LMA) and leaf ash, a proxy for silica—and multiple measures of herbivory by caterpillars of the generalist herbivore fall armyworm (Spodoptera frugiperda). Results: Across all genetic subpopulations, low-latitude plants experienced less herbivory than high-latitude plants. Within genetic subpopulations, however, this relationship was inconsistent—the most widely distributed and phenotypically variable subpopulation (Atlantic) exhibited more consistent latitudinal trends than either of the other two subpopulations. The two physical resistance traits, LMA and leaf ash, were both highly heritable and positively associated with resistance to different measures of herbivory across all subpopulations, indicating their importance in defense against herbivores. Again, however, these relationships were inconsistent within subpopulations. Conclusions: Defensive gradients that occur across the entire species range may not arise within localized subpopulations. Thus, identifying the drivers of latitudinal gradients in herbivory defense may depend on adequately sampling the diversity within a species.

09 BIOMASS FUELS↗

U.S. Department of Energy Free-Air CO 2 Enrichment Experiments: FACE Results, Lessons, and Legacy

In the 1970s, growing concern about potential long-range consequences of carbon dioxide (CO 2 ) emissions from the combustion of fossil fuels prompted the U.S. Department of Energy (DOE) to begin developing a CO 2 research program. The goal was to obtain the scientific information needed to reduce uncertainty and develop a knowledgebase for decision-making. At its start in 1980, this program had three components: the global carbon cycle, climate effects, and vegetation effects. The vegetation effects component recognized that increased atmospheric CO 2 could be a potential benefit to vegetation and crops because CO 2 is an essential resource for plant growth. In addition to increasing crop yield, program developers and researchers recognized that this fertilizer effect could also result in greater storage of fossil fuel CO 2 by forests. Initial experiments were small in scale and scope and focused on the photosynthesis, physiology, and water use of crop plants, but the focus shifted toward responses of plants in natural systems, as needed for connections to the global carbon cycle. Program research investigated CO 2 interactions with other environmental variables, emphasizing field research whenever possible and recognizing that exposure to elevated CO 2 concentrations under realistic field conditions should lead to better predictions of plant response. Whole-ecosystem field experiments were instituted in low-stature ecosystems, but experiments to address forest responses necessarily relied on tree seedlings or saplings in open-top chambers (OTCs) and not within intact forest ecosystems. The need for larger-scale, longer-duration experiments in each of the world’s six major biomes (tundra, boreal forest, temperate forest, tropical forest, grassland, and desert) became clear, and for these experiments, free-air CO 2 exposure facilities were necessary. This report summarizes the objectives and main results of DOE-sponsored experiments employing such facilities, along with the outcomes from research on ecosystem responses using field chambers (see Fig. ES.1, p. x). Additionally, this report highlights the many successes of DOE CO 2 enrichment studies and considers the lessons learned that can be used to guide future initiatives.

54 ENVIRONMENTAL SCIENCES↗

IX Cell Wall Research Conference 2022 (IXCWRC): Cell wall research for new fundamental discoveries and applications in plant biology

This application is for securing funds to offset the registration fees and lodging costs for US-based early career investigators (ERCs) to participate to the IX Cell Wall Research Conference 2022 (IXCWRC 2022): Cell wall research for new fundamental discoveries and applications in plant biology. The conference will be held in East Lansing, Michigan on June 13-17, 2022. The conference will focus on the plant cell wall. Plant cells are encapsulated by the cell wall, a dynamic structure that protects them, enables them to interface with the external environment and supports the growth of the entire organism. There is a growing interest in understanding the complex processes necessary to build and deconstruct the plant cell wall. A better understanding of how the cell wall is synthesized during plant growth and in response to the environment, and how it is deconstructed for a broad variety of applications can inform and improve biotechnological processes for increasing biomass yield and improving industrial processes for the recovery of valuable, sustainable polymers. Acquiring such knowledge requires the exchange of new ideas and results as well as the development and application of new technologies. The IXCWRC 2022 aims to reach this goal by providing an opportunity for an open forum to discuss new ideas and theories and develop collaborations for scientists across the globe in a diverse and supportive environment. A distinction of the IXCWRC 2022 from all the preceding plant cell wall conferences will be its focus on the training and inclusion of the next generation of scientists, the ERCs. The overall goal of this application is to broaden the participation of ERCs with a focus on underrepresented communities. The specific goal of this application is to secure support for registration and lodging for US-based ERCs to attend the conference. Priority will be given to ERCs from underrepresented communities in science. We expect that support from DOE will allow the development of an exciting and innovative program to discuss the latest scientific advances in cell wall biology research, advance the next generation of plant cell wall researchers and foster the foundations of an inclusive community in science.

59 BASIC BIOLOGICAL SCIENCES↗

Earlier Ecological Drought Detection by Involving the Interaction of Phenology and Eco-Physiological Function

Meteorological drought indices like the Standardized Precipitation Evaporation Index (SPEI) are frequently used to diagnose “ecological drought,” despite the fact that they were not explicitly designed for this purpose. More recently developed indices like the Evaporative Stress Index (ESI), which is based on the degree of coupling between actual to potential evapotranspiration, may better capture dynamic plant response to moisture limitations. However, the skill of these indices at describing plant water stress is rarely evaluated at sub-seasonal timescales over which drought evolves. Moreover, it remains unclear how variability in phenological timing impacts and complicates early drought detection. Here, we compared the ability of ESI and SPEI to reflect the dynamics of ecological drought in forests and grasslands, based on anomalies of Gross Primary Productivity (GPP), surface conductance (G s , a proxy for stomatal conductance), soil moisture, and vapor pressure deficit. ESI performed better than SPEI in capturing the dynamics of GPP and G s , but still missed early ecological drought signals due to biases linked to earlier onset of spring leaf development. Thus, we developed a modified variant of the ESI (ESI LAI ) that accounts for the complicating effects of phenological shifts in leaf area index (LAI). The ESI LAI detected drought onset up to 7–10 weeks earlier than SPEI and ESI. Additionally, drought onset dates determined from ESI LAI are close to (±2 weeks) the dates determined from LAI-corrected anomalies of G s , and GPP, as well as the onset dates of soil water deficit and atmospheric aridity.

54 ENVIRONMENTAL SCIENCES↗

Verification testing of the PKI collector at Sandia National Laboratories, Albuquerque, New Mexico

Verification testing of a solar collector was undertaken prior to its operation as part of an industrial process heat plant at Capitol Concrete Products in Topeka, Kansas. Testing was performed at a control plant installed at Sandia National Laboratory, Albuquerque, New Mexico (SNLA). Early results show that plant performance is even better than anticipated and far in excess of test criteria. Overall plant efficiencies of 65 to 80 percent were typical during hours of good insolation. A number of flaws and imperfections were detected during operability testing, the most important being a problem in elevation drive alignment due to a manufacturing error. All problems were corrected as they occurred and the plant, with over 40 hours of operation, is currently continuing operability testing in a wholly-automatic mode.

Hauger, J. S.↗

The nonlinear mechanics of highly extensible plant epidermal cell walls

Plant epidermal cell walls maintain the mechanical integrity of plants and restrict organ growth. Mechanical analyses can give insights into wall structure and are inputs for mechanobiology models of plant growth. To better understand the intrinsic mechanics of epidermal cell walls and how they may accommodate large deformations during growth, we analyzed a geometrically simple material, onion epidermal strips consisting of only the outer (periclinal) cell wall, ~7 μm thick. With uniaxial stretching by >40%, the wall showed complex three-phase stress–strain responses while cyclic stretching revealed reversible and irreversible deformations and elastic hysteresis. Stretching at varying strain rates and temperatures indicated the wall behaved more like a network of flexible cellulose fibers capable of sliding than a viscoelastic composite with pectin viscosity. We developed an analytic framework to quantify nonlinear wall mechanics in terms of stiffness, deformation, and energy dissipation, finding that the wall stretches by combined elastic and plastic deformation without compromising its stiffness. We also analyzed mechanical changes in slightly dehydrated walls. Their extension became stiffer and more irreversible, highlighting the influence of water on cellulose stiffness and sliding. This study offers insights into the structure and deformation modes of primary cell walls and presents a framework that is also applicable to tissues and whole organs.

59 BASIC BIOLOGICAL SCIENCES↗

Efficient delivery of a DNA aptamer-based biosensor into plant cells for glucose sensing through thiol-mediated uptake

DNA aptamers have been widely used as biosensors for detecting a variety of targets. Despite decades of success, they have not been applied to monitor any targets in plants, even though plants are a major platform for providing oxygen, food, and sustainable products ranging from energy fuels to chemicals, and high-value products such as pharmaceuticals. A major barrier to progress is a lack of efficient methods to deliver DNA into plant cells. We herein report a thiol-mediated uptake method that more efficiently delivers DNA into Arabidopsis and tobacco leaf cells than another state-of-the-art method, DNA nanostructures. Such a method allowed efficient delivery of a glucose DNA aptamer sensor into Arabidopsis for sensing glucose. This demonstration opens a new avenue to apply DNA aptamer sensors for functional studies of various targets, including metabolites, plant hormones, metal ions, and proteins in plants for a better understanding of the biodistribution and regulation of these species and their functions.

59 BASIC BIOLOGICAL SCIENCES↗

Data for Efficient Delivery of a DNA Aptamer-Based Biosensor into Plant Cells for Glucose Sensing through Thiol-Mediated Uptake

DNA aptamers have been widely used as biosensors for detecting a variety of targets. Despite decades of success, they have not been applied to monitor any targets in plants, even though plants are a major platform for providing oxygen, food, and sustainable products ranging from energy fuels to chemicals, and high-value products such as pharmaceuticals. A major barrier to progress is a lack of efficient methods to deliver DNA into plant cells. We herein report a thiol-mediated uptake method that more efficiently delivers DNA into Arabidopsis and tobacco leaf cells than another state-of-the-art method, DNA nanostructures. Such a method allowed efficient delivery of a glucose DNA aptamer sensor into Arabidopsis for sensing glucose. This demonstration opens a new avenue to apply DNA aptamer sensors for functional studies of various targets, including metabolites, plant hormones, metal ions, and proteins in plants for a better understanding of the biodistribution and regulation of these species and their functions.

Conversion↗

Time-lapse imagery in 2017 and 2018 at the Lower Montane site in the East River Watershed, Colorado

Time-lapse imagery was collected using an automated RGB camera mounted on a pole at the base of the northeast-facing hillslope at the Lower Montane site in the East River Watershed, Colorado. The imagery was intended to support a better understanding of plant dynamics and their controls during the growing season. The dataset includes RGB images archived in four zip files (containing imagery in JPEG format), corresponding to photos taken from the hillslope and the adjacent floodplain during 2017 and 2018. A fifth zip file contains a few AVI movies that compare imagery between the two years. The AVI files can be read with most media players applications. The archive contains a total of five *.zip files and three csv metadata files (flmd.csv, dd.csv, and locations.csv).This work was supported by the Watershed Function Science Focus Area at Lawrence Berkeley National Laboratory funded by the US Department of Energy, Office of Science, Biological and Environmental Research under Contract No. DE-AC02-05CH11231.

54 ENVIRONMENTAL SCIENCES↗

Structure of Lipopolysaccharide from Liberibacter crescens Is Low Molecular Weight and Offers Insight into Candidatus Liberibacter Biology

Huanglongbing (HLB) disease, also known as citrus greening disease, was first reported in the US in 2005. Since then, the disease has decimated the citrus industry in Florida, resulting in billions of dollars in crop losses and the destruction of thousands of acres of citrus groves. The causative agent of citrus greening disease is the phloem limited pathogen Candidatus Liberibacter asiaticus. As it has not been cultured, very little is known about the structural biology of the organism. Liberibacter are part of the Rhizobiaceae family, which includes nitrogen-fixing symbionts of legumes as well as the Agrobacterium plant pathogens. To better understand the Liberibacter genus, a closely related culturable bacterium (Liberibacter crescens or Lcr) has attracted attention as a model organism for structural and functional genomics of Liberibacters. Given that the structure of lipopolysaccharides (LPS) from Gram-negative bacteria plays a crucial role in mediating host-pathogen interactions, we sought to characterize the LPS from Lcr. We found that the major lipid A component of the LPS consisted of a pentaacylated molecule with a β-6-GlcN disaccharide backbone lacking phosphate. The polysaccharide portion of the LPS was unusual compared to previously described members of the Rhizobiaceae family in that it contained ribofuranosyl residues. The LPS structure presented here allows us to extrapolate known LPS structure/function relationships to members of the Liberibacter genus which cannot yet be cultured. It also offers insights into the biology of the organism and how they manage to effectively attack citrus trees.

59 BASIC BIOLOGICAL SCIENCES↗

Prediction of condition-specific regulatory genes using machine learning

Recent advances in genomic technologies have generated data on large-scale protein–DNA interactions and open chromatin regions for many eukaryotic species. How to identify condition-specific functions of transcription factors using these data has become a major challenge in genomic research. To solve this problem, we have developed a method called ConSReg, which provides a novel approach to integrate regulatory genomic data into predictive machine learning models of key regulatory genes. Using Arabidopsis as a model system, we tested our approach to identify regulatory genes in data sets from single cell gene expression and from abiotic stress treatments. Our results showed that ConSReg accurately predicted transcription factors that regulate differentially expressed genes with an average auROC of 0.84, which is 23.5–25% better than enrichment-based approaches. To further validate the performance of ConSReg, we analyzed an independent data set related to plant nitrogen responses. ConSReg provided better rankings of the correct transcription factors in 61.7% of cases, which is three times better than other plant tools. We applied ConSReg to Arabidopsis single cell RNA-seq data, successfully identifying candidate regulatory genes that control cell wall formation. Our methods provide a new approach to define candidate regulatory genes using integrated genomic data in plants.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Economic viability of using thermal energy storage for flexible carbon capture on natural gas power plants

Fossil fuel-based power plants generate 80% of the electricity in the United States and provide a reliable generation source for both base and peak power demands. These plants are expected to adapt to changes in environmental policies that will require carbon management with carbon capture and storage (CCS) representing a possible solution. Current solvent-based CCS has a detrimental impact on a power plant's performance due to large heat loads required for carbon capture solvent regeneration. This parasitic load restricts the power plant's output and operation flexibility. Therefore, this study evaluates the feasibility of using thermal storage technologies for natural gas combined cycle (NGCC) power plants coupled with CCS to minimize the impact of solvent regeneration and enable the plant to operate at peak power output. Thermal storage can minimize the impact of CCS on the power plant by providing the heat load required for solvent regeneration during times of peak demand which will allow the plant to operate unrestricted and at full power. In total, fifteen unique thermal storage configurations were evaluated from three thermal storage categories: Brayton cycle heat pump, vapor compression heat pump, and heat recovery steam generator steam extraction for storage. The viability of these systems was determined by evaluating each configuration on thousands of real-world Locational Marginal Pricing (LMP) profiles from the New York Independent System Operator and California Independent System Operator electricity markets using a techno-economic analysis. Afterwards, results were compared to the performance of a base power plant (NGCC with CCS and no thermal storage) to determine the impact of thermal storage on power plant economics. Overall, six of the thermal storage configurations performed better than base CCS enabled power plant on between 11.5% and 38.7% of the LMP signals evaluated. The best performing configuration was a vapor compression heat pump that used flue gas as the working fluid and had both hot and cold thermal storage units. This configuration performed better than the base CCS power plant on 38.7% of the LMP profiles. The results of this study show thermal storage can mitigate the economic impact of carbon capture solvent regeneration on NGCC power plants. Discussion focuses on the impact of electricity pricing on the optimal thermal storage system, the advantages and disadvantages of the systems evaluated, and identifies limitations with the study.

25 ENERGY STORAGE↗

A KBase Case Study on Genome-wide Transcriptomics and Plant Primary Metabolism in Sorghum

A better understanding of the genetic and metabolic mechanisms that confer stress resistance and tolerance in plants is key to engineering new crops through advanced breeding technologies. This requires a systems biology approach that builds on a genome-wide understanding of the regulation of gene expression, plant metabolism, physiology and growth. In this study, we examine the response to drought stress in Sorghum, as we leverage the tools for transcriptomics and plant metabolic modeling we have implemented at the U.S. Department of Energy Systems Biology Knowledgebase (KBase). KBase enables researchers worldwide to collaborate and advance research by allowing them to upload private or public data into the KBase Narrative Interface, empowering them to analyze this data using a rich, extensible array of computational and data-analytics tools, and allows them to securely share scientific workflows and conclusions. We demonstrate how to use the current RNA-seq tools in KBase, applicable to both plants and microbes, to assemble and quantify long transcripts and identify differentially expressed genes effectively. More specifically, we demonstrate the utility of the platform by identifying key genes that are differentially expressed during drought-stress in Sorghum bicolor, which is an important sustainable production crop plant. We then show how to use KBase tools to predict the membership of genes in metabolic pathways and examine expression data in the context of metabolic subsystems. We demonstrate the power of the platform by making the data analysis and interpretation available to the biologists in the reproducible, re-usable, point-and-click format of a KBase Narrative thus promoting FAIR (Findable, Accessible, Interoperable and Reusable) guiding principles for scientific data management and stewardship.

59 BASIC BIOLOGICAL SCIENCES↗

Enhancing SWAT with mechanistic plant hydraulics: development and application in the Hanjiang River Basin

Plant transpiration plays a critical role in global water and energy cycles, requiring better process understanding as climate change intensifies drought stress and alters plant responses. Most hydrological models such as the widely-used SWAT lack representation of plant hydraulics, the mechanistic processes controlling plant water regulation and transpiration. Here, this study developed SWAT-PHS by integrating a plant hydraulics scheme (PHS) into SWAT hydrological model, enabling explicit simulation of root water uptake, sap flow, storage and transpiration at 30-minute timescales for watershed-scale modeling. In the Hanjiang River Basin, SWAT-PHS mitigated overestimation of runoff during the rainy season and underestimation during the dry season, reducing the overall simulation error by 29% across the entire simulation period. The model can simulate reasonable plant water dynamics, including diurnal transpiration patterns and drought responses showing declining transpiration flux, hydraulic buffering through stem water storage, and depth-dependent root water uptake strategies. Sensitivity analysis shows that SWAT-PHS captured mechanistic relationships between plant hydraulic traits and transpiration, with root distribution and stem capacitance positively affecting annual transpiration while vulnerability parameters showed negative effects. This work provides a pathway for improving hydrologic modeling and water resource management by better representing plant water regulation under climate change and expected intensifying water stress conditions.

China↗

The complex circuitry of interactions determining coexistence among plants and mycorrhizal fungi

We present a mechanistic model of coexistence among a mycorrhizal fungus and one or two plant species that compete for a single nutrient. Plant–fungal coexistence is more likely if the fungus is better at extracting the environmental nutrient than the plant and the fungus acquires carbon from the plant above a minimum rate. When they coexist, their interaction can shift from mutualistic to parasitic at high nutrient availability. The fungus is a second nutrient source for plants and can promote the coexistence of two plant competitors if one is better at environmental nutrient extraction and the other is better at acquiring the nutrient from the fungus. Because it extracts carbon from both plants, the fungus also serves as a conduit of apparent competition between the plants. Consequently, the plant with the lower environmental nutrient extraction rate can drive the plant with the higher environmental nutrient extraction rate extinct at high carbon supply rates. Furthermore, this model illustrates mechanisms to explain several observed patterns, including shifts in plant–mycorrhizal growth responses and coexistence along nutrient gradients, equivocal results among experiments testing the effect of mycorrhizal fungi on plant diversity, and differences in plant diversity among ecosystems dominated by different mycorrhizal groups.

54 ENVIRONMENTAL SCIENCES↗