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At least 109 records · Page 6

A multi-omic characterization of the physiological responses to salt stress in Scenedesmus obliquus UTEX393

Scenedesmus obliquus UTEX393 is a promising microalgal candidate for sustainable biomanufacturing but its limited halotolerance hinders large-scale cultivation in saline environments. To investigate the molecular basis of salt stress responses, we conducted a comprehensive multi-omic analysis integrating genomics, transcriptomics, proteomics, lipidomics, metabolomics, and DNA affinity purification sequencing (DAP-seq). An improved nuclear genome assembly and annotation yielded 19,017 gene models and a 97% BUSCO completeness score, enabling construction of a genome-scale metabolic model. Comparing 15 ppt salinity stress to 5 ppt control, growth and productivity were significantly reduced, accompanied by widespread transcriptomic and proteomic changes. Transcriptomic analysis revealed downregulation of photosynthetic machinery and energy conservation genes, and upregulation of stress-responsive elements such as expansins, flavodoxins, and osmoprotectants. Lipidomic profiling showed accumulation of triacylglycerols (TAGs) and degradation of galactosyl lipids, consistent with a shift toward lipid biosynthesis to mitigate redox imbalance. Depletion of key polar metabolites and branched-chain amino acids suggested a rerouting of central carbon metabolism under stress. DAP-seq identified key transcription factors, including LHY1 and SPL12, that target central metabolic enzymes involved in redox balancing, such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and malate dehydrogenase (MDH). These findings establish a regulatory-metabolic framework linking redox stress to lipid accumulation and reveal potential engineering targets to enhance salt tolerance. Overall, the multi-omic analysis supports the “overflow” hypothesis, where impaired photosynthesis results in excess reducing equivalents being diverted into TAG synthesis and highlights transcriptional regulators as candidates for improving algal robustness in brackish environments.

09 BIOMASS FUELS↗

Future increases in Amazonia water stress from CO 2 physiology and deforestation

Several different drivers are contributing to climate change within the Amazon basin, including forcing from greenhouse gases and aerosols, plant physiology responses to rising CO 2 , and deforestation. Attribution among these drivers has not been quantified for Shared Socioeconomic Pathway (SSP) climate simulations. Here we identify the contribution of CO 2 physiology and deforestation to future hydroclimate change in the Amazon basin by combining information from four experiments and eight different Earth system models in Coupled Model Intercomparison Project Phase 6. Together, forcing from CO 2 physiology and deforestation account for about 44% of the projected annual precipitation decline, 48% of surface relative humidity decline and 11% of warming over the Amazon basin by 2100 for SSP3-7.0. Further, other Coupled Model Intercomparison Project Phase 6 SSP simulations have similar contributions from the two drivers. Insight from our attribution analysis can aid in identifying research priorities aimed at reducing uncertainty in future projections of water availability, carbon dynamics and wildfire risk.

54 ENVIRONMENTAL SCIENCES↗

Remote sensing of changes in morphology and physiology of trees under stress

Results of continuing studies of forest trees subjected to varying types of stress are reported. Both greenhouse and field studies are included. Greenhouse work with tree seedlings exposed to varying levels of NaCl and CaCl2 in the soil indicated that, in the initial stages, palisade cells shrink and the amount of air space in the leaf increases. As the severity of damage increases, the cells of the spongy mesophyll shrink and flatten, and the amount of air space in the leaf decreases. Statistical analysis of foliar reflectance and associated moisture content data led to a series of regression equations for predicting foliar moisture content from reflectance data. Equations were calculated for three species, yellow birch (Betula alleghaniensis Britton), sugar maple (Acer saccharum Marsh.) and white ash (Fraxinus americana L.) having multiple correlation coefficients of 0.98, 0.94 and 0.93 respectively. Interpretation of multispectral imagery of the Ann Arbor Forestry Test Site (NASA Site 190) provided evidence that infections of Fomes annosus can be detected in the early stages. Infections of two needle cast diseases were also detected in conifer plantations in the test site. A study of automatic interpretation of multispectral scanner imagery for tree species recognition provided encouraging results.

Olson, C. E., Jr.↗

Remote sensing of changes in morphology and physiology of trees under stress

Measurements on foliage samples collected from several drought and salt treated plants revealed that leaf thickness decreased with increasing severity of the drought treatment and increased with increasing severity of treatment with NaCl, but remained essentially unaffected by treatment with CaCl2. Airborne data collected by multispectral scanner indicated that false color images provide selective enhancement of a diseased area. Comparison of simulated and actual aerial color and color IR photography revealed that the color renditions of the MSS simulations agreed closely with those of the actual photography.

Olson, C. E., Jr.↗

Remote sensing of changes in morphology and physiology of trees under stress

Previsual detection of Fomes annosus in pine plantations was studied. Detailed analyses of photographic imagery obtained over the Ann Arbor Test Site during 1969 and 1970 reveal that the Ektachrome Infrared film was superior to Ektachrome Aerographic, Infrared Aerographic, or Plus-X Aerographic films for detecting Fomes annosus damage. Of far more significance in controlling the accuracy of damage detection, however, was the experience of the photo interpreter. Ratio-processing of multispectral scanner data was investigated with data collected in June of 1970 and in June of 1972. Ratioing of the 1.5-1.8 and 1.0-1.4 micrometer channels gave good results at detecting openings in the crown canopy and adjacent infected trees. Combined level slicing of the 1.5-1.8 micrometer channel and the 1.5-1.8 to 1.0-1.4 micrometer ratio permitted separation and recognition of forest litter in the openings and stressed trees adjacent ot the openings.

Olson, C. F., Jr.↗

Gravity and acceleration

Space flight gravity and acceleration effects on human physiological systems - human ecology

ACCELERATION STRESS↗

Ponderosa pine hydraulic stress predicts more extreme wildfire behavior under future conditions in Bandelier National Monument, New Mexico

Background Live fuel moisture contributes to wildfire spread and reflects plant stress and physiological traits. The anticipated change in live fuel moisture under future conditions is likely non-linear, owing to physiological plant thresholds in water hydraulics. We constructed a mechanistic model of live fuel moisture’s response to water stress to understand the impact of future climate on live fuel moisture. We first gathered data on plant physiology and live fuel moisture for Pinus ponderosa at Bandelier National Monument, NM, USA, and modeled their relationship. We then parameterized a mechanistic plant hydrodynamics model (FATES-HYDRO) to simulate changes in plant stress and a statistical model to simulate the resulting impact on live fuel moisture. We then simulated FATES-HYDRO under future climate anomalies (SSP2-4.5 and SSP5-8.5: 2080–2100) to understand the change in plant stress and estimate its impact on live fuel moisture. Results We found that the number of days below crucial thresholds of live fuel moisture (100% and 79%) increased from contemporary levels (< 100%: 72 days, < 79%: 1.4 days) under SSP2-4.5 (< 100%: 185 days, < 79%: 10.2 day) and increased exponentially under SSP5-8.5 (< 100%: 215 days, < 79%: 65 days). We found that gross primary productivity decreased under both future climate scenarios (contemporary: 336 g C m −2 , SSP2-4.5: 203 g C m −2 , SSP5-8.5: 243 g C m −2 ); however, spring productivity increased under SSP5-8.5, potentially altering fuel loading. We additionally see a potentially lethal loss of conductivity in hydraulic P. ponderosa under SSP5-8.5. Conclusions The overall increase in plant water stress (as represented by loss of hydraulic conductivity and leaf water potential) and lower live fuel moisture appear to be driven by reduced precipitation during late summer monsoons typical of the region, extending the fire season later in the year. We predict increasing variability in the P. ponderosa wildfire regime under both climate projections driven by changing productivity, rising mortality, and an overall decrease in live fuel moisture.

54 ENVIRONMENTAL SCIENCES↗

Combined environmental stresses

Tolerance levels, physiological effects, and performance degradation during simultaneous or sequential exposures to two environmental stresses, and also three or more simultaneous stresses are described. Environmental stress combinations are characterized by four descriptors: order of occurrence, duration of exposure, severity of exposure, and type of interaction. Combined stress data and facilities for combined stress study are briefly mentioned.

Murray, R. H.↗

Novel gas exposure system for the controlled exposure of plants to gaseous hydrogen fluoride

Abstract Plants can serve as sensitive bioindicators of the presence of contaminant vapors in the atmosphere. This work describes a novel laboratory-based gas exposure system capable of calibrating plants as bioindicators for the detection and delineation of the atmospheric contaminant hydrogen fluoride (HF) as a preparatory step for monitoring release emissions. To evaluate changes in plant phenotype and stress-induced physiological effects attributed to HF alone, the gas exposure chamber must have additional controls to simulate otherwise optimal plant growth conditions including variables such as light intensity, photoperiod, temperature, and irrigation. The exposure system was designed to maintain constant growth conditions during a series of independent experiments that varied between optimal (control) and stressful (HF exposure) conditions. The system was also designed to ensure the safe handling and application of HF. An initial system calibration introduced HF gas into the exposure chamber and monitored HF concentrations by cavity ring-down spectroscopy for a 48-h period. Stable concentrations inside the exposure chamber were observed after approximately 15 h, and losses of HF to the system ranged from 88 to 91%. A model plant species ( Festuca arundinacea ) was then exposed to HF for 48 h. Visual phenotype stress-induced responses aligned with symptoms reported in the literature for fluoride exposure (tip dieback and discoloration along the dieback transition margin). Fluoride concentrations in exposed tissues compared to control tissues confirmed enhanced fluoride uptake due to HF exposure. The system described herein can be applied to other reactive atmospheric pollutants of interest in support of bioindicator research.

54 ENVIRONMENTAL SCIENCES↗

Effects of Radiation and Dietary Iron on Expression of Genes and Proteins Involved in Drug Metabolism

Liver function, especially the rate of metabolic enzyme activities, determines the concentration of circulating drugs and the duration of their efficacy. Most pharmaceuticals are metabolized by the liver, and clinically-used medication doses are given with normal liver function in mind. A drug overdose can result in the case of a liver that is damaged and removing pharmaceuticals from the circulation at a rate slower than normal. Alternatively, if liver function is elevated and removing drugs from the system more quickly than usual, it would be as if too little drug had been given for effective treatment. Because of the importance of the liver in drug metabolism, we want to understand any effects of spaceflight on the enzymes of the liver. Dietary factors and exposure to radiation are aspects of spaceflight that are potential oxidative stressors and both can be modeled in ground experiments. In this experiment, we examined the effects of high dietary iron and low dose gamma radiation (individually and combined) on the gene expression of enzymes involved in drug metabolism, redox homeostasis, and DNA repair. METHODS All procedures were approved by the JSC Animal Care and Use Committee. Male Sprague-Dawley rats were divided into 4 groups (n=8); control, high Fe diet (650 mg iron/kg), radiation (fractionated 3 Gy exposure from a Cs- 137 source) and combined high Fe diet + radiation exposure. Animals were euthanized 24h after the last treatment of radiation; livers were removed immediately and flash -frozen in liquid nitrogen. Expression of genes thought to be involved in redox homeostasis, drug metabolism and DNA damage repair was measured by RT-qPCR. Where possible, protein expression of the same genes was measured by western blotting. All data are expressed as % change in expression normalized to reference gene expression; comparisons were then made of each treatment group to the sham exposed/ normal diet control group. Data was considered significant at p< 0.5. RESULTS Among the redox homeostasis genes examined, metallothionein showed a significant down regulation in the radiation treated group (-3.85 fold) and a trend toward down regulation in the high Fe + rad group. Metallothionein is involved in the regulation of physiological metals and also has antioxidant activities. Among the drug metabolism genes examined, ATP binding cassette subfamily B (Abcb1b) gene expression increased more than 10-fold in both groups that received radiation treatments. This increased expression was also seen at the protein level. This ABC transporter carries many different compounds across cell membranes, including administered medications. The cytochrome P450 2E1 enzyme, a mixed-function oxidase that deactivates some medications and activates others, showed about a 2-fold increase in gene expression in both radiation-treated groups, with a trend toward increased expression at the protein level. Expression of epoxide hydrolase, which detoxifies polycyclic aromatic hydrocarbons, showed similar 2-fold increases. Among the DNA repair genes examined, expression of RAD51 was significantly down regulated (1.5 fold) in the radiation treated group. RAD51 is involved in repair of double-stranded DNA breaks. CONCLUSION This experiment used 2 different sources of physiological oxidative stress, administered separately and together, and examined their impacts on liver gene and protein expression. It is clear that significant changes occurred in expression of several genes and proteins in the radiation-treated animals. If the results from this ground analog of portions of the spaceflight environment hold true for the spaceflight environment itself, the physiological roles of the affected enzymes (drug transport and metabolism, redox homeostasis) could mean consequences in redox homeostasis or the pharmacokinetics of administered medications

Faust, K. M.↗

Three-dimensional imaging of mitochondrial cristae complexity using cryo-soft X-ray tomography

Mitochondria are dynamic organelles that change morphology to adapt to cellular energetic demands under both physiological and stress conditions. Cardiomyopathies and neuronal disorders are associated with structure-related dysfunction in mitochondria, but three-dimensional characterizations of the organelles are still lacking. In this study, we combined high-resolution imaging and 3D electron density information provided by cryo-soft X-ray tomography to characterize mitochondria cristae morphology isolated from murine. Using the linear attenuation coefficient, the mitochondria were identified (0.247 ± 0.04 µm -1 ) presenting average dimensions of 0.90 ± 0.20 µm in length and 0.63 ± 0.12 µm in width. The internal mitochondria structure was successfully identified by reaching up the limit of spatial resolution of 35 nm. The internal mitochondrial membranes invagination (cristae) complexity was calculated by the mitochondrial complexity index (MCI) providing quantitative and morphological information of mitochondria larger than 0.90 mm in length. The segmentation to visualize the cristae invaginations into the mitochondrial matrix was possible in mitochondria with MCI ≥ 7. Altogether, we demonstrated that the MCI is a valuable quantitative morphological parameter to evaluate cristae modelling and can be applied to compare healthy and disease state associated to mitochondria morphology.

3-D reconstruction↗

Root system architecture and environmental flux analysis in mature crops using 3D root mesocosms

Current methods of root sampling typically only obtain small or incomplete sections of root systems and do not capture their true complexity. To facilitate the visualization and analysis of full-sized plant root systems in 3-dimensions, we developed customized mesocosm growth containers. While highly scalable, the design presented here uses an internal volume of 45 ft 3 (1.27 m 3 ), suitable for large crop and bioenergy grass root systems to grow largely unconstrained. Furthermore, they allow for the excavation and preservation of 3-dimensional root system architecture (RSA), and facilitate the collection of time-resolved subterranean environmental data. Sensor arrays monitoring matric potential, temperature and CO 2 levels are buried in a grid formation at various depths to assess environmental fluxes at regular intervals. Methods of 3D data visualization of fluxes were developed to allow for comparison with root system architectural traits. Following harvest, the recovered root system can be digitally reconstructed in 3D through photogrammetry, which is an inexpensive method requiring only an appropriate studio space and a digital camera. We developed a pipeline to extract features from the 3D point clouds, or from derived skeletons that include point cloud voxel number as a proxy for biomass, total root system length, volume, depth, convex hull volume and solidity as a function of depth. Ground-truthing these features with biomass measurements from manually dissected root systems showed a high correlation. We evaluated switchgrass, maize, and sorghum root systems to highlight the capability for species wide comparisons. We focused on two switchgrass ecotypes, upland (VS16) and lowland (WBC3), in identical environments to demonstrate widely different root system architectures that may be indicative of core differences in their rhizoeconomic foraging strategies. Finally, we imposed a strong physiological water stress and manipulated the growth medium to demonstrate whole root system plasticity in response to environmental stimuli. Hence, these new “3D Root Mesocosms” and accompanying computational analysis provides a new paradigm for study of mature crop systems and the environmental fluxes that shape them.

59 BASIC BIOLOGICAL SCIENCES↗