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At least 37 records · Page 2

Microbial Mutualism in (Modeled) Microgravity: Measuring the Effects of A Spaceflight-Like Fluid Environment on Cross-Feeding Communities

Microorganisms grow differently in spaceflight than they do on Earth. While numerous factors likely contribute, one ubiquitous feature of the space environment is altered gravity - but because microbes are generally considered too small to detect gravity directly, they likely experience the effects of gravity via changes in their fluid environment, primarily a reduction in convective mixing. This theory is supported by evidence that individual microbial strains experience starvation and acid stress in microgravity. If it is true, then microgravity would also have measurable effects on microbes in mixed communities, because many interspecies interactions involve the exchange of soluble metabolites through their medium (cross-feeding). Specifically, cooperative cross-feeding communities would grow more slowly in microgravity, and this effect should be reproducible in ground-based rotational culture systems (rotating wall vessels, RWVs) designed to simulate the quiescent environment of microgravity. Here we present preliminary results of our efforts to test this hypothesis using using a model system of Eschericia coli and Salmonella enterica that grow only when they can exchange methionine and acetate through the medium. We report on the development of a novel laboratory system that allows us measure growth rates and species ratios in real-time using fluorescence spectrometry as the organisms grow in RWVs. We are able to track growth rates of individual species and the co-culture continuously with high temporal resolution for several hours, a substantial improvement over prior RWV systems that require destructive sampling. We vary the fidelity of the microgravity simulation by varying the rotation rate, and find that rotation rate affects growth rates of the community more than that of individual strains, consistent with the claim that RWVs can simulate a microgravity-like quiescent environment. While more work needs to be done to probe the mechanisms of this effect, we hope that our system will ultimately allow us to generate quantitative predictions about the effects of spaceflight on organisms that are tiny but essential to sustaining long-term human space exploration.

microgravity↗

Effects of mechanical stress or abscisic acid on growth, water status and leaf abscisic acid content of eggplant seedlings

Container-grown eggplant (Solanum melongena L. var esculentum Nees. 'Burpee's Black Beauty') seedlings were conditioned with brief, periodic mechanical stress or abscisic acid (ABA) in a greenhouse prior to outdoor exposure. Mechanical stress consisted of seismic (shaking) or thigmic (stem flexing) treatment. Exogenous ABA (10(-3) or 10(-4)M) was applied as a soil drench 3 days prior to outdoor transfer. During conditioning, only thigmic stress reduced stem elongation and only 10(-3) M ABA reduced relative growth rate (RGR). Both conditioning treatments increased leaf specific chlorophyll content, but mechanical stress did not affect leaf ABA content. Outdoor exposure of unconditioned eggplant seedlings decreased RGR and leaf-specific chlorophyll content, but tended to increase leaf ABA content relative to that of plants maintained in the greenhouse. Conditioning did not affect RGR of plants subsequently transferred outdoors, but did reduce stem growth. Seismic stress applied in the greenhouse reduced dry weight gain by plants subsequently transferred outdoors. Mechanical stress treatments increased leaf water potential by 18-25% relative to that of untreated plants.

NASA Discipline Plant Biology↗

Osmotic stress, endogenous abscisic acid and the control of leaf morphology in Hippuris vulgaris L

Previous reports indicate that heterophyllous aquatic plants can be induced to form aerial-type leaves on submerged shoots when they are grown in exogenous abscisic acid (ABA). This study reports on the relationship between osmotic stress (e.g. the situation encountered by a shoot tip when it grows above the water surface), endogenous ABA (as measured by gas chromatography-electron capture detector) and leaf morphology in the heterophyllous aquatic plant, Hippuris vulgaris. Free ABA could not be detected in submerged shoots of H. vulgaris but in aerial shoots ABA occurred at ca. 40 ng (g fr wt)-1. When submerged shoots were osmotically stressed ABA appeared at levels of 26 to 40 ng (g fr wt)-1. These and other data support two main conclusions: (1) Osmotically stressing a submerged shoot causes the appearance of detectable levels of ABA. (2) The rise of ABA in osmotically stressed submerged shoots in turn induces a change in leaf morphology from the submerged to the aerial form. This corroborates the hypothesis that, in the natural environment, ABA levels rise in response to the osmotic stress encountered when a submerged shoot grows up through the water/air interface and that the increased ABA leads to the production of aerial-type leaves.

NASA Program Space Biology↗

Metabolomic profiles are reflective of hypoxia-induced insulin resistance during exercise in healthy young adult males

Hypoxia-induced insulin resistance appears to suppress exogenous glucose oxidation during metabolically matched aerobic exercise during acute (<8 h) high-altitude (HA) exposure. However, a better understanding of this metabolic dysregulation is needed to identify interventions to mitigate these effects. The objective of this study was to determine if differences in metabolomic profiles during exercise at sea level (SL) and HA are reflective of hypoxia-induced insulin resistance. Native lowlanders ( n = 8 males) consumed 145 g (1.8 g/min) of glucose while performing 80-min of metabolically matched treadmill exercise at SL (757 mmHg) and HA (460 mmHg) after 5-h exposure. Exogenous glucose oxidation and glucose turnover were determined using indirect calorimetry and dual tracer technique ([ 13 C]glucose and [6,6- 2 H 2 ]glucose). Metabolite profiles were analyzed in serum as change (Δ), calculated by subtracting postprandial/exercised state SL (ΔSL) and HA (ΔHA) from fasted, rested conditions at SL. Compared with SL, exogenous glucose oxidation, glucose rate of disappearance, and glucose metabolic clearance rate (MCR) were lower ( P < 0.05) during exercise at HA. One hundred and eighteen metabolites differed between ΔSL and ΔHA ( P < 0.05, Q < 0.10). Differences in metabolites indicated increased glycolysis, tricarboxylic acid cycle, amino acid catabolism, oxidative stress, and fatty acid storage, and decreased fatty acid mobilization for ΔHA. Branched-chain amino acids and oxidative stress metabolites, Δ3-methyl-2-oxobutyrate ( r = -0.738) and Δγ-glutamylalanine ( r = -0.810), were inversely associated ( P < 0.05) with Δexogenous glucose oxidation. Δ3-Hydroxyisobutyrate ( r = -0.762) and Δ2-hydroxybutyrate/2-hydroxyisobutyrate ( r = -0.738) were inversely associated ( P < 0.05) with glucose MCR. Coupling global metabolomics and glucose kinetic data suggest that the underlying cause for diminished exogenous glucose oxidative capacity during aerobic exercise is acute hypoxia-mediated peripheral insulin resistance.

59 BASIC BIOLOGICAL SCIENCES↗

The effect of fluid mechanical stress on cellular arachidonic acid metabolism

The effect of sublytic levels of mechanical perturations of cells on cell metabolism were investigated by analyzing the products of arachidonic acid (used as a marker metabolite) in blood platelets, polymorphonuclear leucocytes, and cultured umbilical-vein endothelial cells after the suspensions of these cells were subjected to a shear stress in a modified viscometer. It is shown that the sublytic levels of mechanical stress stimulated the arachidonic acid metabolism in all these cell types. Possible biological implications of this stress-metabolism coupling are discussed.

Mcintire, L. V.↗

Spaceflight-Induced Changes in Microbial Virulence and the Impact to the Host Immune Response

Many microbial pathogen shave repeatedly exhibited unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in final cell concentration, biofilm production, stress resistance, antibiotic sensitivity, gene expression, host-pathogen interactions, and virulence. Notably, the classic foodborne pathogen Salmonella enterica serovar Typhimurium displayed increased virulence in animals when cultured in either the spaceflight analogue or true spaceflight environment. Recently, Serratia marcescens also was shown to increase virulence when cultured in the spaceflight environment. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system at multiple levels, which suggests an increased risk of infectious diseases. Moreover, astronauts have some degree of clinical infectious disease incidence. However, the contribution of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. The goal of this study is to gain insight into the breadth of other medically significant microbial pathogens that may exhibit altered virulence and pathogenesis-related responses when cultured in space flight analogue conditions. Specifically, we are characterizing the effect of spaceflight analogue culture (Low Shear Modeled Microgravity/LSMMG) on microbial pathogenesis-related stress responses, in vitro host-pathogen interactions, gene expression, and virulence potential in animals using five important model bacterial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli. Herein, we present data from one of these pathogens, the foodborne bacterium, S. enterica Enteritidis, which is closely related to S. enterica Typhimurium. Phenotypes evaluated included growth profiles, environmental stress responses(acid, oxidative, bile, and thermal stresses),and in vitro colonization of3-D biomimetic cultures of human intestinal tissue containing immune cells. Transcriptomic profiling and virulence studies are ongoing. We show that S. Enteritidis exhibited key alterations in pathogenic responses to LSMMG culture that suggest increased infection risk, including several responses which were different from those observed in the closely related pathovar S. Typhimurium. This information will provide critical mechanistic insight into the potential impact of microgravity on alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.

C M Ott↗

Spaceflight-Induced Changes in Microbial Virulence and the Impact to the Host Immune Response

Many microbial pathogen shave repeatedly exhibited unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in final cell concentration, biofilm production, stress resistance, antibiotic sensitivity, gene expression, host-pathogen interactions, and virulence. Notably, the classic foodborne pathogen Salmonella enterica serovar Typhimurium displayed increased virulence in animals when cultured in either the spaceflight analogue or true spaceflight environment. Recently, Serratia marcescens also was shown to increase virulence when cultured in the spaceflight environment. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system at multiple levels, which suggests an increased risk of infectious diseases. Moreover, astronauts have some degree of clinical infectious disease incidence. However, the contribution of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. The goal of this study is to gain insight into the breadth of other medically significant microbial pathogens that may exhibit altered virulence and pathogenesis-related responses when cultured in space flight analogue conditions. Specifically, we are characterizing the effect of spaceflight analogue culture (Low Shear Modeled Microgravity/LSMMG) on microbial pathogenesis-related stress responses, in vitro host-pathogen interactions, gene expression, and virulence potential in animals using five important model bacterial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli. Herein, we present data from one of these pathogens, the foodborne bacterium, S. enterica Enteritidis, which is closely related to S. enterica Typhimurium. Phenotypes evaluated included growth profiles, environmental stress responses(acid, oxidative, bile, and thermal stresses),and in vitro colonization of3-D biomimetic cultures of human intestinal tissue containing immune cells. Transcriptomic profiling and virulence studies are ongoing. We show that S. Enteritidis exhibited key alterations in pathogenic responses to LSMMG culture that suggest increased infection risk, including several responses which were different from those observed in the closely related pathovar S. Typhimurium. This information will provide critical mechanistic insight into the potential impact of microgravity on alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.

C.M. Ott↗

Decreases in polyunsaturated fatty acid content improve heat stress tolerance during flowering and silicle development in pennycress (Thlaspi arvense L.)

Introduction: Pennycress (Thlaspi arvense L.) is an emerging intermediate oilseed crop grown in the offseason between primary summer crops to produce three cash crops in two years. Previous efforts to improve seed oil quality produced Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) genome-edited lines with decreased polyunsaturated fatty acids (PUFAs) levels, through loss of function of the FATTY ACID DESATURASE 2 (FAD2), REDUCED OLEATE DESATURATION1 (ROD1), and FATTY ACID ELONGASE1 (FAE1) genes. While seed oil compositions were previously characterized, it remains unknown how vegetative and reproductive tissue compositions might differ and affect tolerance to high temperature (HT) conditions.Methods: In four growth chamber experiments, we explored HT tolerance during flowering and silicle development. Plants were subjected to a 34 °C day/28 °C night regime and compared to control plants maintained at 20 °C. Pollen grain viability at a range of temperatures, lipid peroxidation and proline content in leaves and silicles following HT, and seed yield were measured.Results: Both fad2 and rod1 mutant lines had relatively higher pollen viability (71% and 54% respectively) under moderately elevated temperature (28 °C) compared to wild-type controls (37%). They also showed smaller decreases in seed yield (0% and 40% for fad2 and rod1 respectively, compared to 61% for wild type), following HT exposure during late flowering and early silicle development. Silicles of fad2 plants experienced 65% less lipid peroxidation under HT and 55% less buildup of proline, signifying less stress.Discussion: The differential results of fad2 and rod1 are likely due to the role of FAD2 in membranes in all tissues, whereas ROD1 predominantly affects triacylglycerol (TAG) composition in oil-accumulating tissues including pollen. Our results indicate that decreasing PUFAs, through gene editing, can increase heat tolerance in reproductive tissues as an auxiliary benefit accompanying improved seed oil quality.

59 BASIC BIOLOGICAL SCIENCES↗

The ulcerogenic effect of bile and bile acid in rats during immobilization stress

The effect of different concentrations of oxen bile and individual bile acids or their sodium salts on the gastric mucosa of rats was investigated in combination with immobilization stress. A statistically significant higher frequency of ulcers was only determined in the application of 10% oxen bile. Dosages on 10% sodium glycocholic acid demonstrated strong toxic damage with atonic dilation of the stomach and extensive mucosal bleeding.

Weisener, J.↗

Formation, Survival and Evolution of Organics After Impact Processing of Asteroidal and Cometary Simulants

This work examines the formation and evolution of amino acids during impact events on the prebiotic Earth Laboratory experiments, which simulate impact processes, provide direct tests of questions regarding the conditions (concentration of amino acid, shock stress or impact velocity, matrix material) under which amino acid synthesis and polymerization occur.

exobiology↗

Engineering robust microorganisms for organic acid production

Abstract Organic acids are an important class of compounds that can be produced by microbial conversion of renewable feedstocks and have huge demands and broad applications in food, chemical, and pharmaceutical industries. An economically viable fermentation process for production of organic acids requires robust microbial cell factories with excellent tolerance to low pH conditions, high concentrations of organic acids, and lignocellulosic inhibitors. In this review, we summarize various strategies to engineer robust microorganisms for organic acid production and highlight their applications in a few recent examples.

59 BASIC BIOLOGICAL SCIENCES↗

The stimulation of arachidonic acid metabolism in human platelets by hydrodynamic stresses

The effects of stimulating human platelets by thrombin and by hydrodynamic stresses on the platelets' arachidonic acid metabolism were investigated using (1-C-14)-arachidonic acid label and a specially designed viscometer that ensured laminar shear flow with a nearly uniform shear rate throughout the flow region. It was found that platelets activated by thrombin formed principally thromboxane A2, 12-hydroxy 5,8,10-heptadecatrienoic acid and 12-hydroxy 5,8,10,14-eicosatetraenoic acid (12-HETE). On the other hand, platelets activated by shear, formed only 12-HETE (although arachidonic acid metabolism was stimulated); no cyclooxygenase metabolites were detected. Results indicate that platelets may greatly increase their 12-HETE production when activated by passage through a high-stress region of the circulation, such as an atherosclerotic stenosis.

Rajagopalan, Sridhar↗

Spatiotemporal metabolic responses to water deficit stress in distinct leaf cell-types of poplar

The impact of water-deficit (WD) stress on plant metabolism has been predominantly studied at the whole tissue level. However, plant tissues are made of several distinct cell types with unique and differentiated functions, which limits whole tissue ‘omics’-based studies to determine only an averaged molecular signature arising from multiple cell types. Advancements in spatial omics technologies provide an opportunity to understand the molecular mechanisms underlying plant responses to WD stress at distinct cell-type levels. Here, we studied the spatiotemporal metabolic responses of two poplar ( Populus tremula× P. alba ) leaf cell types -palisade and vascular cells- to WD stress using matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI). We identified unique WD stress-mediated metabolic shifts in each leaf cell type when exposed to early and prolonged WD stresses and recovery from stress. During water-limited conditions, flavonoids and phenolic metabolites were exclusively accumulated in leaf palisade cells. However, vascular cells mainly accumulated sugars and fatty acids during stress and recovery conditions, respectively, highlighting the functional divergence of leaf cell types in response to WD stress. By comparing our MALDI-MSI metabolic data with whole leaf tissue gas chromatography-mass spectrometry (GC-MS)-based metabolic profile, we identified only a few metabolites including monosaccharides, hexose phosphates, and palmitic acid that showed a similar accumulation trend at both cell-type and whole leaf tissue levels. Overall, this work highlights the potential of the MSI approach to complement the whole tissue-based metabolomics techniques and provides a novel spatiotemporal understanding of plant metabolic responses to WD stress. This will help engineer specific metabolic pathways at a cellular level in strategic perennial trees like poplars to help withstand future aberrations in environmental conditions and to increase bioenergy sustainability.

59 BASIC BIOLOGICAL SCIENCES↗

Extreme Low-Temperature Stress Affects Nutritional Quality of Amino Acids in Rice

Global climate change has increased the frequency of extreme climate events, and their effects on the nutritional quality, especially on amino acids in rice, have not been quantified. The data from a 3-year low temperature stress (LTS) experiment including two rice varieties (Huaidao 5 and Nanjing 46), seven minimum/maximum temperature levels (one optimal 21/27°C and six LTS levels from 17/23 to 6/12°C), and three LTS durations (3, 6, and 9 days) after flowering, revealed significant interactive effects of LTS at different stages, durations, and temperature levels on the content and accumulation of amino acids. LTS increased rice total amino acid content, while decreasing its accumulation, with higher sensitivities to LTS at the flowering stage than at the grain filling stage. In most treatments, the lysine (the first limiting amino acid) and phenylalanine content were increased under LTS at early and peak flowering stages but decreased at the grain filling stage in both varieties, and only leucine content was increased at all three stages after flowering, while the content of other essential amino acids differed among the two varieties. With an increase of 1°C·d per day in the accumulated cold degree days, the relative content of the essential amino acids was increased by 0.01–0.41%, depending on the rice variety and growth stage. Our results suggest that LTS can improve nutritional quality of amino acids of rice grains in terms of amino acids content, especially at flowering stage. These results provide critical insights for assessing the potential impact of extreme climates on the nutrient quality of rice under future climate change.

Kang, Min↗

QTL Mapping of Seed Fatty Acid Contents in Camelina sativa Under Heat Stress

Heat stress alters oil quality in oilseed crops, yet its genetic underpinnings in Camelina sativa remain unclear. This study investigated the genetic basis of heat-induced changes in seed fatty acids using a recombinant inbred line (RIL) population derived from a cross between two camelina varieties, Suneson and Pryzeth. Exposure to high temperature during reproductive growth led to increased proportions of saturated (C16:0, C18:0) and monounsaturated (C18:1) fatty acids, whereas polyunsaturated C18:3, total unsaturated fatty acids (UFA) and the PUFA/MUFA ratio were decreased, suggesting an inhibition of the C18:1 → C18:2 → C18:3 desaturation pathway. A high-density linkage map (4981 bins across 20 chromosomes) was built, and 25 QTLs for fatty acids were detected, with hotspots on chromosomes 1, 9, 12, 13, 16, and 20. A major QTL on chromosome 1 (~ 80 cM) explained the largest variance component for PUFA/MUFA under heat. Three desaturase genes (FAD2, FAD7, FAD8) were located within key QTL intervals, nominating them as candidates for modulating unsaturation under elevated temperature. These results provide a genetic basis for fine mapping and functional validation, supporting future molecular and breeding efforts to stabilize oil quality under warming conditions.

Camelina↗

A multi-omic characterization of temperature stress in a halotolerant Scenedesmus strain for algal biotechnology

Microalgae efficiently convert sunlight into lipids and carbohydrates, offering bio-based alternatives for energy and chemical production. Improving algal productivity and robustness against abiotic stress requires a systems level characterization enabled by functional genomics. Here, we characterize a halotolerant microalga Scenedesmus sp. NREL 46B-D3 demonstrating peak growth near 25 °C that reaches 30 g/m 2 /day and the highest biomass accumulation capacity post cell division reported to date for a halotolerant strain. Functional genomics analysis revealed that genes involved in lipid production, ion channels and antiporters are expanded and expressed. Exposure to temperature stress shifts fatty acid metabolism and increases amino acids synthesis. Co-expression analysis shows that many fatty acid biosynthesis genes are overexpressed with specific transcription factors under cold stress. These and other genes involved in the metabolic and regulatory response to temperature stress can be further explored for strain improvement.

09 BIOMASS FUELS↗