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At least 199 records · Page 11
All sourcesNASA NTRSDOE OSTI
DOE OSTI2022

Is Genetic Engineering a Route to Enhance Microalgae-Mediated Bioremediation of Heavy Metal-Containing Effluents?

Contamination of the biosphere by heavy metals has been rising, due to accelerated anthropogenic activities, and is nowadays, a matter of serious global concern. Removal of such inorganic pollutants from aquatic environments via biological processes has earned great popularity, for its cost-effectiveness and high efficiency, compared to conventional physicochemical methods. Among candidate organisms, microalgae offer several competitive advantages; phycoremediation has even been claimed as the next generation of wastewater treatment technologies. Furthermore, integration of microalgae-mediated wastewater treatment and bioenergy production adds favorably to the economic feasibility of the former process—with energy security coming along with environmental sustainability. However, poor biomass productivity under abiotic stress conditions has hindered the large-scale deployment of microalgae. Recent advances encompassing molecular tools for genome editing, together with the advent of multiomics technologies and computational approaches, have permitted the design of tailor-made microalgal cell factories, which encompass multiple beneficial traits, while circumventing those associated with the bioaccumulation of unfavorable chemicals. Previous studies unfolded several routes through which genetic engineering-mediated improvements appear feasible (encompassing sequestration/uptake capacity and specificity for heavy metals); they can be categorized as metal transportation, chelation, or biotransformation, with regulation of metal- and oxidative stress response, as well as cell surface engineering playing a crucial role therein. This review covers the state-of-the-art metal stress mitigation mechanisms prevalent in microalgae, and discusses putative and tested metabolic engineering approaches, aimed at further improvement of those biological processes. Finally, current research gaps and future prospects arising from use of transgenic microalgae for heavy metal phycoremediation are reviewed.

Ranjbar, Saeed (ORCID:0000000256518605)↗
DOE OSTI2024

Comprehensive Genome-Wide Natural Variation and Expression Analysis of Tubby-like Proteins Gene Family in Brachypodium distachyon

The Tubby-like proteins (TLPs) gene family is a group of transcription factors found in both animals and plants. In this study, we identified twelve B. distachyon TLPs, divided into six groups based on conserved domains and evolutionary relationships. We predicted cis-regulatory elements involved in light, hormone, and biotic and abiotic stresses. The expression patterns in response to light and hormones revealed that BdTLP3, 4, 7, and 14 are involved in light responses, and BdTLP1 is involved in ABA responses. Furthermore, BdTLP2, 7, 9, and 13 are expressed throughout vegetative and reproductive stages, whereas BdTLP1, 3, 5, and 14 are expressed at germinating grains and early vegetative development, and BdTLP4, 6, 8, and 10 are expressed at the early reproduction stage. The natural variation in the eleven most diverged B. distachyon lines revealed high conservation levels of BdTLP1-6 to high variation in BdTLP7-14 proteins. Based on diversifying selection, we identified amino acids in BdTLP1, 3, 8, and 13, potentially substantially affecting protein functions. This analysis provided valuable information for further functional studies to understand the regulation, pathways involved, and mechanism of BdTLPs.

Plant Sciences↗
DOE OSTI2025

Plant Defense Proteins: Recent Discoveries and Applications

Proteins play pivotal roles in safeguarding plants against numerous biotic and abiotic stresses. Understanding their biological functions and mechanisms of action is essential for advancing plant biology, agriculture, and biotechnology. This review considers the diversity and potential applications of plant defense proteins including pathogenesis-related (PR) proteins, chitinases, glucanases, protease inhibitors, lectins, and antimicrobial peptides. Recent advances, such as the omics technologies, have enabled the discovery of new plant defense proteins and regulatory networks that govern plant defense responses and unveiled numerous roles of plant defense proteins in stress perception, signal transduction, and immune priming. The molecular affinities and enzymatic activities of plant defense proteins are essential for their defense functions. Applications of plant defense proteins span agriculture, biotechnology, and medicine, including the development of resistant crop varieties, bio-based products, biopharmaceuticals, and functional foods. Future research directions include elucidating the structural bases of defense protein functions, exploring protein interactions with ligands and other proteins, and engineering defense proteins for enhanced efficacy. Overall, this review illuminates the significance of plant defense proteins against biotic stresses in plant biology and biotechnology, emphasizing their potential for sustainable agriculture and environmental management.

G-proteins↗
NASA NTRS2002

Hydrogen peroxide homeostasis: activation of plant catalase by calcium/calmodulin

Environmental stimuli such as UV, pathogen attack, and gravity can induce rapid changes in hydrogen peroxide (H(2)O(2)) levels, leading to a variety of physiological responses in plants. Catalase, which is involved in the degradation of H(2)O(2) into water and oxygen, is the major H(2)O(2)-scavenging enzyme in all aerobic organisms. A close interaction exists between intracellular H(2)O(2) and cytosolic calcium in response to biotic and abiotic stresses. Studies indicate that an increase in cytosolic calcium boosts the generation of H(2)O(2). Here we report that calmodulin (CaM), a ubiquitous calcium-binding protein, binds to and activates some plant catalases in the presence of calcium, but calcium/CaM does not have any effect on bacterial, fungal, bovine, or human catalase. These results document that calcium/CaM can down-regulate H(2)O(2) levels in plants by stimulating the catalytic activity of plant catalase. Furthermore, these results provide evidence indicating that calcium has dual functions in regulating H(2)O(2) homeostasis, which in turn influences redox signaling in response to environmental signals in plants.

NASA Discipline Plant Biology↗
NASA NTRS2004

Ethylene synthesis and sensitivity in crop plants

Closed and semi-closed plant growth chambers have long been used in studies of plant and crop physiology. These studies include the measurement of photosynthesis and transpiration via photosynthetic gas exchange. Unfortunately, other gaseous products of plant metabolism can accumulate in these chambers and cause artifacts in the measurements. The most important of these gaseous byproducts is the plant hormone ethylene (C2H4). In spite of hundreds of manuscripts on ethylene, we still have a limited understanding of the synthesis rates throughout the plant life cycle. We also have a poor understanding of the sensitivity of intact, rapidly growing plants to ethylene. We know ethylene synthesis and sensitivity are influenced by both biotic and abiotic stresses, but such whole plant responses have not been accurately quantified. Here we present an overview of basic studies on ethylene synthesis and sensitivity.

NASA Program Advanced Life Support↗
NASA NTRSDate not supplied

Determining Chlamydomonas reinhardtii resistance to ionizing radiation at a genome-wide scale

Spaceflight is a complex mixture of abiotic stresses that causes a myriad of cellular responses including increased genome-wide mutation load. The impact of spaceflight on mutation load is not well understood for most organisms and is especially critical to characterize for candidate microorganisms that could be part of biological life support systems. Prior research in our laboratory revealed an unexpectedly high level of mutation during spaceflight in the eukaryotic green algae, C. reinhardtii. Based on the spectrum of base changes observed, we concluded that the mutations were generated through a molecular mechanism unique to the spaceflight environment. Spaceflight is a complex set of stresses, and we are using an Earth-based analog to test ionizing radiation as a potential causative variable for the unique mutation spectrum observed in spaceflight. C. reinhardtii was grown exposed to chronic radiation from a Cesium-137 source. Four biological replicate cultures of wild-type cells were grown in gas permeable plastic tissue culture bags in liquid media following the design used on the International Space Station (ISS) during the previous space algae experiment. Radiation levels for this experiment approximate ionizing radiation observed on the moon, in low Earth orbit (LEO), and on Earth. Cells from each replicate were passaged to inoculate the next cycle of growth, then samples were taken from each culture to characterize cell morphology, quantify biomass, and extract DNA. Whole genome sequencing from each culture will be used to identify the frequency and types of mutations in each radiation level. In growth cycles 3 and 4, there was a significant increase in biomass accumulation under lunar radiation level compared to LEO or Earth radiation levels. These results such that ionizing radiation affected growth of C. reinhardtii.

Microbiology↗
NASA NTRSDate not supplied

RNASeq and Fluorescence Analysis of the Response of ERF2 and ERF104 in Arabidopsis thaliana under Simulated Altered Gravity

As NASA moves closer to long-term human space exploration, the need to understand how to sustain life in space is increasingly pressing. Plants are essential to human sustenance, making it important to understand how spaceflight affects plant health. We used differential gene expression analysis to examine GLDS-251 (RNAseq analysis of the response of Arabidopsis thaliana to fractional gravity under blue-light stimulation during spaceflight) from NASA’s GeneLab data repository and found downregulation of ERF2 and ERF104, transcription factors of the ethylene response factor families, that integrate hormonal pathways involved in abiotic stress responses. Downregulation of ERF2 and ERF104 during spaceflight may indicate a dysregulation of the ethylene signaling pathway. Our hypothesis is that altered gravity downregulates the expression of ERF2 and ERF104 in Arabidopsis thaliana, altering the ethylene signaling pathway and affecting the electron transport chain and light-dependent reactions in chloroplast thylakoids. To test this hypothesis, we propose to grow A. thaliana seedlings (wild-type and mutant/knockout of ERF2 and ERF104) in altered gravity conditions to determine the effects on the expression of ERF2, ERF104, and photosynthesis. We anticipate that ERF2 and ERF104 will be underexpressed in altered gravity conditions and result in decreased regulation of the ethylene signaling pathway.

Arabidopsis↗
NASA NTRSDate not supplied

Determining Chlamydomonas Reinhardtii Resistance to Ionizing Radiation at A Genome-Wide Scale

Spaceflight is a complex mixture of abiotic stresses that causes a myriad of cellular responses including increased genome-wide mutation load. The impact of spaceflight on mutation load is not well understood for most organisms and is especially critical to characterize for candidate microorganisms that could be part of biological life support systems. Prior research in our laboratory revealed an unexpectedly high level of mutation during spaceflight in the eukaryotic green algae, C. reinhardtii. Based on the spectrum of base changes observed, we concluded that the mutations were generated through a molecular mechanism unique to the spaceflight environment. Spaceflight is a complex set of stresses, and we are using an Earth-based analog to test ionizing radiation as a potential causative variable for the unique mutation spectrum observed in spaceflight. C. reinhardtii was grown exposed to chronic radiation from a Cesium-137 source. Four biological replicate cultures of wild-type cells were grown in gas permeable plastic tissue culture bags in liquid media following the design used on the International Space Station (ISS) during the previous space algae experiment. Radiation levels for this experiment approximate ionizing radiation observed on the moon, in low Earth orbit (LEO), and on Earth. Cells from each replicate were passaged to inoculate the next cycle of growth, then samples were taken from each culture to characterize cell morphology, quantify biomass, and extract DNA. Whole genome sequencing from each culture will be used to identify the frequency and types of mutations in each radiation level. In growth cycles 3 and 4, there was a significant increase in biomass accumulation under lunar radiation level compared to LEO or Earth radiation levels. These results show that ionizing radiation affected growth of C. reinhardtii.

Hope Hersh↗
NASA NTRS2023

Anomalous Net Biome Exchange Over Amazonian Rainforests Induced by the 2015/16 El Niño: Temperature-dominated Total Flux but Soil Dryness-Shaped Spatial Pattern

The magnitude and spatial pattern of anomalous net biome exchange (NBE) induced by the 2015/16 El Niño over Amazonian rainforests remain uncertain. We here investigated them using multi-model posterior NBE products in the Orbiting Carbon Observatory-2 (OCO-2) version 10 modeling intercomparison project. Results suggest that relative to the annual NBE average in 2017/18, larger anomalous carbon release occurred over the eastern and northern Amazonian rainforests in 2015/16, with a total flux of approximately 0.4 PgC yr−1 after assimilating satellite-observed column CO2 concentrations (XCO2) over land. We further find that this anomalous spatial pattern was predominantly determined by soil dryness, while the total positive NBE anomaly was dominated by higher temperature with its contribution of approximately 68～70%. We believe that atmospheric inversions assimilating more satellite-observed XCO2 in future can provide us more comprehensive understanding how Amazonian rainforests cope with the abiotic stresses induced by El Niño events.

Jun Wang↗
DOE OSTI2025

Breeding of microbiomes conferring salt tolerance to plants

Microbiome breeding through host-mediated selection is a technique to artificially select for microbiomes conferring beneficial properties to plants. Using a systematic selection protocol that maximises the heritability of microbiome effects, transmission fidelity, and microbiome stability through multiple selection cycles, we previously developed root-associated microbial communities conferring sodium and aluminium tolerance to Brachypodium distachyon, a model for cereal crops. Here, we explore the physiological mechanisms underlying our selected microbiomes’ effect on plant fitness and analyse how our selection protocol shaped the composition and structure of these microbiomes. We analysed the effects of our selected microbiomes on plant fitness and tissue-nutrient concentration, then used 16S rRNA amplicon sequencing to examine microbial community composition and co-occurrence network patterns. Our sodium-selected microbiomes reduced leaf sodium concentration by ~ 50%, whereas the aluminium-selected microbiomes had no effect on leaf-tissue nutrient concentration, suggesting different mechanisms underlying the microbiome-mediated stress tolerance. By testing the selected microbiomes in a cross-fostering experiment, we show that our artificially selected microbiomes attained (a) ecological robustness contributing to transplantability (i.e. inheritance) of microbiome-encoded effects between plants; and (b) network features identifying key bacteria promoting salt-stress tolerance. Combined, these findings elucidate critical mechanisms underlying host-mediated artificial selection as a framework to breed microbiomes with targeted benefits for plants under salt stresses, with significant implications for sustainable agriculture.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Responses of the Plant Cell Wall to Sub-Zero Temperatures: A Brief Update

Our general understanding of plant responses to sub-zero temperatures focuses on mechanisms that mitigate stress to the plasma membrane. The plant cell wall receives comparatively less attention, and questions surrounding its role in mitigating freezing injury remain unresolved. Despite recent molecular discoveries that provide insight into acclimation responses, the goal of reducing freezing injury in herbaceous and woody crops remains elusive. This is likely due to the complexity associated with adaptations to low temperatures. Understanding how leaf cell walls of herbaceous annuals promote tissue tolerance to ice does not necessarily lead to understanding how meristematic tissues are protected from freezing by tissue-level barriers formed by cell walls in overwintering tree buds. In this mini-review, we provide an overview of biological ice nucleation and explain how plants control the spatiotemporal location of ice formation. We discuss how sugars and pectin side chains alleviate adhesive injury that develops at sub-zero temperatures between the matrix polysaccharides and ice. The importance of site-specific cell-wall elasticity to promote tissue expansion for ice accommodation and control of porosity to impede ice growth and promote supercooling will be presented. How specific cold-induced proteins modify plant cell walls to mitigate freezing injury will also be discussed. The opinions presented in this report emphasize the importance of a plant’s developmental physiology when characterizing mechanisms of freezing survival.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2026

Data for A Role for Differential Rubisco Activase Isoform Expression in C4 Bioenergy Grasses at High Temperature

Rubisco activase (Rca) facilitates the release of sugar-phosphate inhibitors at Rubisco catalytic sites during CO2 fixation. Most plant species express two Rca isoforms, the larger Rca-α and the shorter Rca-β, either by alternative splicing from a single gene or expression from separate genes. The mechanism of Rubisco activation by Rca isoforms has been intensively studied in C3 plants. However, the functional role of Rca in C4 plants where Rubisco and Rca are located in a much higher [CO2] compartment is less clear. In this study, we selected four C4 bioenergy grasses and the model C4 grass setaria ( Setaria viridis ) to investigate the role of Rca in C4 photosynthesis. All five C4 grass species contained two Rca genes, one encoding Rca-α and the other Rca-β, which were positioned closely together in the genomes. A variety of abiotic stress-related motifs were identified in the Rca-α promoter of each grass, and while the Rca-β gene was constantly highly expressed at ambient temperature, Rca-α isoforms were expressed only at high temperature but never surpassed 30% of Rca-β content. The pattern of Rca-α induction on transition to high temperature and reduction on return to ambient temperature was the same in all five C4 grasses. In sorghum ( Sorghum bicolor ), sugarcane ( Saccharum officinarum ), and setaria, the induction rate of Rca-α was similar to the recovery rate of photosynthesis and Rubisco activation at high temperature. This association between Rca-α isoform expression and maintenance of Rubisco activation at high temperature suggests that Rca-α has a functional thermo-protective role in carbon fixation in C4 grasses by sustaining Rubisco activation at high temperature.

Genomics↗
DOE OSTI2026

Data for "Genetics of flooding tolerance in an F2 Miscanthus sacchariflorus ssp. lutarioriparius × M. sinensis population"

This dataset contains all data and supplementary materials from "Genetics of flooding tolerance in an F2 Miscanthus sacchariflorus ssp. lutarioriparius × M. sinensis population". 1. The dataset S1 table contains the raw phenotypic data collected during the experiment. 2. The dataset S2 table contains the LSmean values for the 24 traits studied. 3. The dataset S3 table contains the TASSEL GBSv2 map, marker information, and genotype data used for mapping. 4. The dataset S4 table contains information on candidate genes found in each of the QTL intervals. 5. The dataset S5 table contains the GO annotations and KEGG enrichment analyses for those candidate genes. 6. The dataset S6 table contains information on the sequences used to classify AP2 ERF transcription factors. 7. The dataset S7 table contains information on AP2 ERF orthologs between Miscanthus and rice based on synteny. 8. Supplementary file 1 contains the ANOVA results using the raw phenotypic data collected from protocol "A". 9. Supplementary file 2 contains the ANOVA results using the raw phenotypic data collected from protocol "B". 10. Supplementary file 3 contains notes on the comparison of SNP calling methods. 11. Supplementary file 4 is a script for analyzing candidate genes found in QTL intervals.

Miscanthus, flood, partial submergence, complete s↗
DOE OSTI2023

Stressing the importance of plant specialized metabolites: omics-based approaches for discovering specialized metabolism in plant stress responses

Plants produce a diverse range of specialized metabolites that play pivotal roles in mediating environmental interactions and stress adaptation. These unique chemical compounds also hold significant agricultural, medicinal, and industrial values. Despite the expanding knowledge of their functions in plant stress interactions, understanding the intricate biosynthetic pathways of these natural products remains challenging due to gene and pathway redundancy, multifunctionality of proteins, and the activity of enzymes with broad substrate specificity. In the past decade, substantial progress in genomics, transcriptomics, metabolomics, and proteomics has made the exploration of plant specialized metabolism more feasible than ever before. Notably, recent advances in integrative multi-omics and computational approaches, along with other technologies, are accelerating the discovery of plant specialized metabolism. In this review, we present a summary of the recent progress in the discovery of plant stress-related specialized metabolites. Emphasis is placed on the application of advanced omics-based approaches and other techniques in studying plant stress-related specialized metabolism. Additionally, we discuss the high-throughput methods for gene functional characterization. These advances hold great promise for harnessing the potential of specialized metabolites to enhance plant stress resilience in the future.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRSDate not supplied

Engineering 6-Phosphogluconate Improves Grain Yield in Heat-Stressed Maize

Endosperm starch synthesis is a primary determinant of grain yield and is sensitive to high-temperature stress. The maize chloroplast-localized 6-phosphogluconate dehydrogenase (6PGDH), PGD3, is critical for endosperm starch accumulation. Maize also has two cytosolic isozymes, PGD1 and PGD2, that are not required for kernel development. We found that cytosolic PGD1 and PGD2 isozymes have heat-stable activity, while amyloplast-localized PGD3 activity is labile under heat stress conditions. We targeted heat-stable 6PGDH to endosperm amyloplasts by fusing the Waxy1 chloroplast targeting the peptide coding sequence to the Pgd1 and Pgd2 open reading frames (ORFs). These WPGD1 and WPGD2 fusion proteins import into isolated chloroplasts, demonstrating a functional targeting sequence. Transgenic maize plants expressing WPGD1 and WPGD2 with an endosperm-specific promoter increased 6PGDH activity with enhanced heat stability in vitro. WPGD1 and WPGD2 transgenes complement the pgd3-defective kernel phenotype, indicating the fusion proteins are targeted to the amyloplast. In the field, the WPGD1 and WPGD2 transgenes can mitigate grain yield losses in high-nighttime-temperature conditions by increasing kernel number. These results provide insight into the subcellular distribution of metabolic activities in the endosperm and suggest the amyloplast pentose phosphate pathway is a heat-sensitive step in maize kernel metabolism that contributes to yield loss during heat stress.

Synthetic Biology↗
DOE OSTI2021

The core autophagy machinery is not required for chloroplast singlet oxygen-mediated cell death in the Arabidopsis thaliana plastid ferrochelatase two mutant

Chloroplasts respond to stress and changes in the environment by producing reactive oxygen species (ROS) that have specific signaling abilities. The ROS singlet oxygen ( 1 O 2 ) is unique in that it can signal to initiate cellular degradation including the selective degradation of damaged chloroplasts. This chloroplast quality control pathway can be monitored in the Arabidopsis thaliana mutant plastid ferrochelatase two ( fc2 ) that conditionally accumulates chloroplast 1 O 2 under diurnal light cycling conditions leading to rapid chloroplast degradation and eventual cell death. The cellular machinery involved in such degradation, however, remains unknown. Recently, it was demonstrated that whole damaged chloroplasts can be transported to the central vacuole via a process requiring autophagosomes and core components of the autophagy machinery. The relationship between this process, referred to as chlorophagy, and the degradation of 1 O 2 -stressed chloroplasts and cells has remained unexplored. Results To further understand 1 O 2 -induced cellular degradation and determine what role autophagy may play, the expression of autophagy-related genes was monitored in 1 O 2 -stressed fc2 seedlings and found to be induced. Although autophagosomes were present in fc2 cells, they did not associate with chloroplasts during 1 O 2 stress. Mutations affecting the core autophagy machinery ( atg5 , atg7 , and atg10 ) were unable to suppress 1 O 2 -induced cell death or chloroplast protrusion into the central vacuole, suggesting autophagosome formation is dispensable for such 1 O 2 –mediated cellular degradation. However, both atg5 and atg7 led to specific defects in chloroplast ultrastructure and photosynthetic efficiencies, suggesting core autophagy machinery is involved in protecting chloroplasts from photo-oxidative damage. Finally, genes predicted to be involved in microautophagy were shown to be induced in stressed fc2 seedlings, indicating a possible role for an alternate form of autophagy in the dismantling of 1 O 2 -damaged chloroplasts. Conclusions Our results support the hypothesis that 1 O 2 -dependent cell death is independent from autophagosome formation, canonical autophagy, and chlorophagy. Furthermore, autophagosome-independent microautophagy may be involved in degrading 1 O 2 -damaged chloroplasts. At the same time, canonical autophagy may still play a role in protecting chloroplasts from 1 O 2 -induced photo-oxidative stress. Together, this suggests chloroplast function and degradation is a complex process utilizing multiple autophagy and degradation machineries, possibly depending on the type of stress or damage incurred.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Multiple pathways mediate chloroplast singlet oxygen stress signaling

Chloroplasts can respond to stress and changes in the environment by producing reactive oxygen species (ROS). Aside from being cytotoxic, ROS also have signaling capabilities. For example, the ROS singlet oxygen ( 1 O 2 ) can initiate nuclear gene expression, chloroplast degradation, and cell death. Here to unveil the signaling mechanisms involved, researchers have used several 1 O 2 -producing Arabidopsis thaliana mutants as genetic model systems, including plastid ferrochelatase two (fc2), fluorescent in blue light (flu), chlorina 1 (ch1), and accelerated cell death 2 (acd2). Here, we compare these 1 O 2 -producing mutants to elucidate if they utilize one or more signaling pathways to control cell death and nuclear gene expression. Using publicly available transcriptomic data, we demonstrate fc2, flu, and ch1 share a core response to 1 O 2 accumulation, but maintain unique responses, potentially tailored to respond to their specific stresses. Subsequently, we used a genetic approach to determine if these mutants share 1 O 2 signaling pathways by testing the ability of genetic suppressors of one 1 O 2 producing mutant to suppress signaling in a different 1 O 2 producing mutant. Our genetic analyses revealed at least two different chloroplast 1 O 2 signaling pathways control cellular degradation: one specific to the flu mutant and one shared by fc2, ch1, and acd2 mutants, but with life-stage-specific (seedling vs. adult) features. Overall, this work reveals chloroplast stress signaling involving 1 O 2 is complex and may allow cells to finely tune their physiology to environmental inputs.

59 BASIC BIOLOGICAL SCIENCES↗