Search NASA⌕ Search

SEARCH · Search NASA

Results for “Root Cause Analysis”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 307 records · Page 17
All sourcesNASA NTRSDOE OSTI
NASA NTRS1984

Measurement of thigmomorphogenesis and gravitropism by non-intrusive computerized video image processing

A video image processing instrument, DARWIN (Digital Analyser of Resolvable Whole-pictures by Image Numeration), was developed. It was programmed to measure stem or root growth and bending, and coupled to a specially mounted video camera to be able to automatically generate growth and bending curves during gravitropism. The growth of the plant is recorded on a video casette recorder with a specially modified time lapse function. At the end of the experiment, DARWIN analyses the growth or movement and prints out bending and growth curves. This system was used to measure thigmomorphagenesis in light grown corn plants. If the plant is rubbed with an applied force load of 0.38 N., it grows faster than the unrubbed control, whereas 1.14 N. retards its growth. Image analysis shows that most of the change in the rate of growth is caused in the first hour after rubbing. When DARWIN was used to measure gravitropism in dark grown oat seedlings, it was found that the top side of the shoot contracts during the first hour of gravitational stimulus, whereas the bottom side begins to elongate after 10 to 15 minutes.

Jaffe, M. J.↗
DOE OSTI2023

Low-pass spectral analysis of time-resolved serial femtosecond crystallography data

Low-pass spectral analysis (LPSA) is a recently developed dynamics retrieval algorithm showing excellent retrieval properties when applied to model data affected by extreme incompleteness and stochastic weighting. In this work, we apply LPSA to an experimental time-resolved serial femtosecond crystallography (TR-SFX) dataset from the membrane protein bacteriorhodopsin (bR) and analyze its parametric sensitivity. While most dynamical modes are contaminated by nonphysical high-frequency features, we identify two dominant modes, which are little affected by spurious frequencies. The dynamics retrieved using these modes shows an isomerization signal compatible with previous findings. We employ synthetic data with increasing timing uncertainty, increasing incompleteness level, pixel-dependent incompleteness, and photon counting errors to investigate the root cause of the high-frequency contamination of our TR-SFX modes. By testing a range of methods, we show that timing errors comparable to the dynamical periods to be retrieved produce a smearing of dynamical features, hampering dynamics retrieval, but with no introduction of spurious components in the solution, when convergence criteria are met. Using model data, we are able to attribute the high-frequency contamination of low-order dynamical modes to the high levels of noise present in the data. Finally, we propose a method to handle missing observations that produces a substantial dynamics retrieval improvement from synthetic data with a significant static component. Reprocessing of the bR TR-SFX data using the improved method yields dynamical movies with strong isomerization signals compatible with previous findings.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Are Atmospheric Models Too Cold in the Mountains? The State of Science and Insights from the SAIL Field Campaign

Mountains play an outsized role in water resource availability, and the amount and timing of water they provide depend strongly on temperature. To that end, we ask the question: How well are atmospheric models capturing mountain temperatures? We synthesize results showing that high-resolution, regionally relevant climate models produce 2-m air temperature (T2m) measurements colder than what is observed (a “cold bias”), particularly in snow-covered midlatitude mountain ranges during winter. We find common cold biases in 44 studies across global mountain ranges, including single-model and multimodel ensembles. We explore the factors driving these biases and examine the physical mechanisms, data limitations, and observational uncertainties behind T2m. Our analysis suggests that the biases are genuine and not due to observation sparsity or resolution mismatches. Cold biases occur primarily on mountain peaks and ridges, whereas valleys are often warm biased. Our literature review suggests that increasing model resolution does not clearly mitigate the bias. By analyzing data from the Surface Atmosphere Integrated Field Laboratory (SAIL) field campaign in the Colorado Rocky Mountains, we test various hypotheses related to cold biases and find that local wind circulations, longwave (LW) radiation, and surface-layer parameterizations contribute to the T2m biases in this particular location. We conclude by emphasizing the value of coordinated model evaluation and development efforts in heavily instrumented mountain locations for addressing the root cause(s) of T2m biases and improving predictive understanding of mountain climates.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2024

Are Atmospheric Models Too Cold in the Mountains? The State of Science and Insights from the SAIL Field Campaign

Mountains play an outsized role in water resource availability, and the amount and timing of water they provide depend strongly on temperature. To that end, we ask the question: How well are atmospheric models capturing mountain temperatures? We synthesize results showing that high-resolution, regionally relevant climate models produce 2-m air temperature (T2m) measurements colder than what is observed (a “cold bias”), particularly in snow-covered midlatitude mountain ranges during winter. We find common cold biases in 44 studies across global mountain ranges, including single-model and multimodel ensembles. We explore the factors driving these biases and examine the physical mechanisms, data limitations, and observational uncertainties behind T2m. Our analysis suggests that the biases are genuine and not due to observation sparsity or resolution mismatches. Cold biases occur primarily on mountain peaks and ridges, whereas valleys are often warm biased. Our literature review suggests that increasing model resolution does not clearly mitigate the bias. By analyzing data from the Surface Atmosphere Integrated Field Laboratory (SAIL) field campaign in the Colorado Rocky Mountains, we test various hypotheses related to cold biases and find that local wind circulations, longwave (LW) radiation, and surface-layer parameterizations contribute to the T2m biases in this particular location. We conclude by emphasizing the value of coordinated model evaluation and development efforts in heavily instrumented mountain locations for addressing the root cause(s) of T2m biases and improving predictive understanding of mountain climates.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2025

Disruption of the endogenous indole glucosinolate pathway impacts the Arabidopsis thaliana root exudation profile and rhizobacterial community

Root exudates are composed of primary and secondary metabolites known to modulate the rhizosphere microbiota. Glucosinolates are defense compounds present in the Brassicaceae family capable of deterring pathogens, herbivores and biotic stressors in the phyllosphere. In addition, traces of glucosinolates and their hydrolyzed byproducts have been found in the soil, suggesting that these secondary metabolites could play a role in the modulation and establishment of the rhizosphere microbial community associated with this family. Here, we used Arabidopsis thaliana mutant lines, including the cyp79B2cyp79B3 double mutant line with a disruption in the indole glucosinolate pathway and atr1D, which overexpresses ATR1 and increases glucosinolate production. These lines were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and 16S rRNA amplicon sequencing to evaluate how genetic modifications to the indole glucosinolate pathway affects the root exudate profile of Arabidopsis thaliana, and, in turn, impacts the rhizosphere microbial community. Metabolic analysis of root exudates from the wild-type Columbia (Col-0), along with the mutant lines, confirmed that alterations to the indole glucosinolate biosynthetic pathway result in shifts in the root exudate profile of the plant. We observed changes in the relative abundance of exuded metabolites. Moreover, 16S rRNA amplicon sequencing results provided evidence that the rhizobacterial communities associated with the plant lines used were directly impacted in diversity and community composition. Here, this work provides further information on the involvement of secondary metabolites and their role in modulating the rhizobacterial community. Root metabolites dictate the presence of different bacterial species, including plant growth-promoting rhizobacteria (PGPR). Our results suggest that genetic alterations in the indole glucosinolate pathway cause disruptions beyond the endogenous levels of the plant, significantly changing the abundance and presence of different metabolites in the root exudates of the plants as well as the microbial rhizosphere community.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS2017

Mass and Reliability Source (MaRS) Database

The Mass and Reliability Source (MaRS) Database consolidates components mass and reliability data for all Oribital Replacement Units (ORU) on the International Space Station (ISS) into a single database. It was created to help engineers develop a parametric model that relates hardware mass and reliability. MaRS supplies relevant failure data at the lowest possible component level while providing support for risk, reliability, and logistics analysis. Random-failure data is usually linked to the ORU assembly. MaRS uses this data to identify and display the lowest possible component failure level. As seen in Figure 1, the failure point is identified to the lowest level: Component 2.1. This is useful for efficient planning of spare supplies, supporting long duration crewed missions, allowing quicker trade studies, and streamlining diagnostic processes. MaRS is composed of information from various databases: MADS (operating hours), VMDB (indentured part lists), and ISS PART (failure data). This information is organized in Microsoft Excel and accessed through a program made in Microsoft Access (Figure 2). The focus of the Fall 2017 internship tour was to identify the components that were the root cause of failure from the given random-failure data, develop a taxonomy for the database, and attach material headings to the component list. Secondary objectives included verifying the integrity of the data in MaRS, eliminating any part discrepancies, and generating documentation for future reference. Due to the nature of the random-failure data, data mining had to be done manually without the assistance of an automated program to ensure positive identification.

Valdenegro, Wladimir↗
DOE OSTI2025

Symbiosis Under Stress: How Arbuscular Mycorrhizal Fungi and Sorghum Metabolism Shift Under Drought

As drought becomes more prevalent across the globe, causing billions of dollars in agricultural loss, the need to maintain crop health and productivity grows increasingly important. Out of the most important cereal crops, sorghum shows the greatest drought tolerance, and plant-microbiome interactions at the root region play a crucial role in this. A key microbial player is arbuscular mycorrhizal fungi (AMF), which deliver water and nutrients to plants in exchange for nutrients they cannot produce. Using sorghum as a model plant, we combine mass-spectrometry based proteomics and metabolomics to examine metabolic interactions between sorghum roots and AMF under drought stress. With AMF, sorghum downregulated lipid-related biological processes involving fatty acid biosynthesis and metabolism. Similarly, amino acid biosynthesis pathways were also suppressed; however, metabolite analysis revealed increased amino acid abundance related to the phenylpropanoid pathway. AMF also enhanced the upregulation of drought-protective osmolytes, such as mannitol and sorbitol, suggesting their key role in mediating sorghum’s response to drought stress. AMF hyphal biomass also had an increased abundance of key osmoprotectant amino acids, indicating similar mechanisms of drought tolerance between sorghum and AMF. Metabolomic evidence also suggested that carbohydrate exchange between sorghum and AMF shifted under stress, indicating an altered exudation pattern likely driven by drought response. Our results demonstrate the molecular mechanisms through which AMF modulate sorghum metabolism under drought conditions, highlighting their promising role in improving crop resilience. By identifying the molecular targets that can improve drought tolerance, we can begin engineering drought resistant agricultural biosystems.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2023

A Divergent Clade KAI2 Protein in the Root Parasitic Plant Orobanche minor Is a Highly Sensitive Strigolactone Receptor and Is Involved in the Perception of Sesquiterpene Lactones

Abstract Strigolactones (SLs) were initially discovered as germination inducers for root parasitic plants. In 2015, three groups independently reported the characterization of the SL receptor in the root parasitic plant Striga hermonthica, which causes significant damage to crop production, particularly in sub-Saharan Africa. The characterized receptors belong to HYPOSENSITIVE TO LIGHT/KARRIKIN INSENSITIVE2 (HTL/KAI2), which is a member of the α/β-hydrolase protein superfamily. In non-parasitic plants, HTL/KAI2 perceives the smoke-derived germination inducer karrikin and a yet-unidentified endogenous ligand. However, root parasitic plants evolved a specific clade of HTL/KAI2 that has diverged from the KAI2 clade of non-parasitic plants. The S. hermonthica SL receptors are included in this specific clade, which is called KAI2 divergent (KAI2d). Orobanche minor is an obligate root holoparasitic plant that grows completely dependent on the host for water and nutrients because of a lack of photosynthetic ability. Previous phylogenetic analysis of KAI2 proteins in O. minor has demonstrated the presence of at least five KAI2d clade genes. Here, we report that KAI2d3 and KAI2d4 in O. minor have the ability to act as the SL receptors. They directly interact with SLs in vitro, and when expressed in Arabidopsis, they rescue thermo-inhibited germination in response to the synthetic SL analog GR24. In particular, KAI2d3 showed high sensitivity to GR24 when expressed in Arabidopsis, suggesting that this receptor enables highly sensitive SL recognition in O. minor. Furthermore, we provide evidence that these KAI2d receptors are involved in the perception of sesquiterpene lactones, non-strigolactone-type germination inducers.

Cell Biology↗
NASA NTRS2016

Post-Test Inspection of NASA's Evolutionary Xenon Thruster Long-Duration Test Hardware: Discharge and Neutralizer Cathodes

The NEXT Long-Duration Test is part of a comprehensive thruster service life assessment intended to demonstrate overall throughput capability, validate service life models, quantify wear rates as a function of time and operating condition, and identify any unknown life-limiting mechanisms. The test was voluntarily terminated in February 2014 after demonstrating 51,184 hours of high-voltage operation, 918 kg of propellant throughput, and 35.5 MN-s of total impulse. The post-test inspection of the thruster hardware began shortly afterwards with a combination of non-destructive and destructive analysis techniques, and is presently nearing completion. This paper presents relevant results of the post-test inspection for both discharge and neutralizer cathodes. Discharge keeper erosion was found to be significantly reduced from what was observed in the NEXT 2 kh wear test and NSTAR Extended Life Test, providing adequate protection of vital cathode components throughout the test with ample lifetime remaining. The area of the discharge cathode orifice plate that was exposed by the keeper orifice exhibited net erosion, leading to cathode plate material building up in the cathode-keeper gap and causing a thermally-induced electrical short observed during the test. Significant erosion of the neutralizer cathode orifice was also found and is believed to be the root cause of an observed loss in flow margin. Deposition within the neutralizer keeper orifice as well as on the downstream surface was thicker than expected, potentially resulting in a facility-induced impact on the measured flow margin from plume mode. Neutralizer keeper wall erosion on the beam side was found to be significantly lower compared to the NEXT 2 kh wear test, likely due to the reduction in beam extraction diameter of the ion optics that resulted in decreased ion impingement. Results from the post-test inspection have led to some minor thruster design improvements.

Electric Propulsion↗
NASA NTRS2021

Post-test Inspection of NASA’s Evolutionary Xenon Thruster Long-Duration Test Hardware: Discharge and Neutralizer Cathodes

The NEXT Long-Duration Test is part of a comprehensive thruster service life assessment intended to demonstrate overall throughput capability, validate service life models, quantify wear rates as a function of time and operating condition, and identify any unknown life-limiting mechanisms. The test was voluntarily terminated in February 2014 after demonstrating 51,184 h of high-voltage operation, 918 kg of propellant throughput, and 35.5 MN-s of total impulse. The post-test inspection of the thruster hardware began shortly afterwards with a combination of non-destructive and destructive analysis techniques, and is presently nearing completion. This paper presents relevant results of the post-test inspection for both discharge and neutralizer cathodes. Discharge keeper erosion was found to be significantly reduced from what was observed in the NEXT 2 kh wear test and NSTAR Extended Life Test, providing adequate protection of vital cathode components throughout the test with ample lifetime remaining. The area of the discharge cathode orifice plate that was exposed by the keeper orifice exhibited net erosion, leading to cathode plate material building up in the cathode-keeper gap and causing a thermally-induced electrical short observed during the test. Significant erosion of the neutralizer cathode orifice was also found and is believed to be the root cause of an observed loss in flow margin. Deposition within the neutralizer keeper orifice as well as on the downstream surface was thicker than expected, potentially resulting in a facility-induced impact on the measured flow margin from plume mode. Neutralizer keeper wall erosion on the beam side was found to be significantly lower compared to the NEXT 2 kh wear test, likely due to the reduction in beam extraction diameter of the ion optics that resulted in decreased ion impingement. Results from the post-test inspection have led to some minor thruster design improvements.

Ion engine↗
NASA NTRS2002

Spatial separation of light perception and growth response in maize root phototropism

Although the effects of gravity on root growth are well known and interactions between light and gravity have been reported, details of root phototropic responses are less documented. We used high-resolution image analysis to study phototropism in primary roots of Zea mays L. Similar to the location of perception in gravitropism, the perception of light was localized in the root cap. Phototropic curvature away from the light, on the other hand, developed in the central elongation zone, more basal than the site of initiation of gravitropic curvature. The phototropic curvature saturated at approximately 10 micromoles m-2 s-1 blue light with a peak curvature of 29 +/- 4 degrees, in part due to induction of positive gravitropism following displacement of the root tip from vertical during negative phototropism. However, at higher fluence rates, development of phototropic curvature is arrested even if gravitropism is avoided by maintaining the root cap vertically using a rotating feedback system. Thus continuous illumination can cause adaptation in the signalling pathway of the phototropic response in roots.

NASA Discipline Plant Biology↗
NASA NTRS2012

Soldering in a Reduced Gravity Environment (SoRGE)

Future long-duration human exploration missions will be challenged by constraints on mass and volume allocations available for spare parts. Addressing this challenge will be critical to the success of these missions. As a result, it is necessary to consider new approaches to spacecraft maintenance and repair that reduce the need for large replacement components. Currently, crew members on the International Space Station (ISS) recover from faults by removing and replacing, using backup systems, or living without the function of Orbital Replacement Units (ORUs). These ORUs are returned to a depot where the root cause of the failure is determined and the ORU is repaired. The crew has some limited repair capability with the Modulation/DeModulation (MDM) ORU, where circuit cards are removed and replace in faulty units. The next step to reducing the size of the items being replaced would be to implement component-level repair. This mode of repair has been implemented by the U.S. Navy in an operational environment and is now part of their standard approach for maintenance. It is appropriate to consider whether this approach can be adapted for future spaceflight operations. To this end, the Soldering in a Reduced Gravity Environment (SoRGE) experiment studied the effect of gravity on the formation of solder joints on electronic circuit boards. This document describes the SoRGE experiment, the analysis methods, and results to date. This document will also contain comments from the crew regarding their experience conducting the SoRGE experiment as well as recommendations for future improvements. Finally, this document will discuss the plans for the SoRGE samples which remain on ISS.

Easton, John W.↗
NASA NTRS2012

STS-133 Space Shuttle External Tank Intertank Stringer Crack Investigation Stress Analysis

The first attempt to launch the STS-133 Space Shuttle mission in the fall of 2010 was halted due to indications of a gaseous hydrogen leak at the External Tank ground umbilical carrier plate seal. Subsequent inspection of the external tank (figure 1) hardware and recorded video footage revealed that the foam insulation covering the forward end of the intertank near the liquid oxygen tank had cracked severely enough to have been cause for halting the launch attempt on its own (figure 2). An investigation into the cause of the insulation crack revealed that two adjacent hat-section sheet metal stringers (figure 3) had cracks up to nine inches long in the forward ends of the stringer flanges, or feet, near the fasteners that attach the stringer to the skin of the intertank (figure 4). A repair of those two stringers was implemented and the investigation effort widened to understand the root cause of the stringer cracks and to determine whether there was sufficient flight rationale to launch with the repairs and the other installed stringers.

Steeve, Brian E.↗
NASA NTRS1994

Detection of atmospheric pressure loading using very long baseline interferometry measurements

Loading of the Earth by the temporal redistribution of global atmospheric mass is likely to displace the positions of geodetic monuments by tens of millimeters both vertically and horizontally. Estimates of these displacements are determined by convolving National Meteorological Center (NMC) global values of atmospheric surface pressure with Farrell's elastic Green's functions. An analysis of the distances between radio telescopes determined by very long baseline interferometry (VLBI) between 1984 and 1992 reveals that in many of the cases studied there is a significant contribution to baseline length change due to atmospheric pressure loading. Our analysis covers intersite distances of between 1000 and 10,000 km and is restricted to those baselines measured more than 100 times. Accounting for the load effects (after first removing a best fit slope) reduces the weighted root-mean-square (WRMS) scatter of the baseline length residuals on 11 of the 22 baselines investigated. The slight degradation observed in the WRMS scatter on the remaining baselines is largely consistent with the expected statistical fluctuations when a small correction is applied to a data set having a much larger random noise. The results from all baselines are consistent with approximately 60% of the computed pressure contribution being present in the VLBI length determinations. Site dependent coefficients determined by fitting local pressure to the theoretical radial displacement are found to reproduce the deformation caused by the regional pressure to within 25% for most inland sites. The coefficients are less reliable at near coastal and island stations.

Vandam, T. M.↗
DOE OSTI2022

Application of Orthogonal Defect Classification for Software Reliability Analysis

The modernization of existing and new nuclear power plants with digital instrumentation and control systems (DI&C) is a recent and highly trending topic. However, there lacks strong consensus on best-estimate reliability methodologies by both the United States (U.S.) Nuclear Regulatory Commission (NRC) and the industry. This has resulted in hesitation for further modernization projects until a more unified methodology is realized. In this work, we develop an approach called Orthogonal-defect Classification for Assessing Software Reliability (ORCAS) to quantify probabilities of various software failure modes in a DI&C system. The method utilizes accepted industry methodologies for software quality assurance that are also verified by experimental or mathematical formulations. In essence, the approach combines a semantic failure classification model with a reliability growth model to predict (and quantify) the potential failure modes of a DI&C software system. The semantic classification model is used to address the question: How do latent defects in software contribute to different software failure root causes? The use of reliability growth models is then used to address the question: Given the connection between latent defects and software failure root causes, how can we quantify the reliability of the software? A case study was conducted on a representative I&C platform (ChibiOS) running a smart sensor acquisition software developed by Virginia Commonwealth University (VCU). The testing and evidence collection guidance in ORCAS was applied, and defects were uncovered in the software. Qualitative evidence, such as condition coverage, was used to gauge the completeness and trustworthiness of the assessment while quantitative evidence was used to determine the software failure probabilities. The reliability of the software was then estimated and compared to existing operational data of the sensor device. It is demonstrated that by using ORCAS, a semantic reasoning framework can be developed to justify if the software is reliable (or unreliable) while still leveraging the strength of the existing methods.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗
NASA NTRSDate not supplied

Expanding the Spatiotemporal Range of Soil Moisture Analysis Utilizing NASA Earth Observations and In Situ Measurements

Drought can cause immense agricultural and ecological damage resulting in high mitigation and compensation costs. Climate variability in future decades is expected to cause severe drought conditions and threaten necessary water resources. Stakeholders seek to implement effective drought assessments in preparation for potential economic and environmental damage invoked by drought. Although in-situ measurements are accurate, the current infrastructure is spatially limited and costly to maintain. A framework was created to compare modeled, satellite and in-situ data in drought monitoring. Here we show that the comparison of in-situ and remotely sensed soil moisture (SM) measurements can increase the spatiotemporal range of SM assessments. Data collected between 2003 and 2021 by NASA’s SPoRT Land Information System (SPoRT-LIS) and Soil Moisture Active Passive (SMAP) mission were standardized and compared with in-situ data provided through the Illinois Climate Network (WARM). Statistical analysis results including the Pearson correlation coefficient (r), root mean squared error, mean absolute error and others were calculated to compare the WARM measurements to the SMAP and SPoRT-LIS products. Results indicate that both satellite products demonstrate seasonally variable bias that is not present in the in-situ measurements. Bias was highest in the winter months and lowest in the late summer and early fall months in both satellite datasets. Overall, WARM-SPoRT comparisons resulted in lower seasonal variability. However, on average, the SMAP comparison demonstrated higher correlation values and lower error values. The WARM-SMAP average correlation (r) was 0.61 compared to the WARM-SPoRT average correlation (r) value of 0.54. Average mean absolute error values calculated for the SMAP and SPoRT comparisons were 0.07 and 0.08 percent soil moisture by volume, respectively. These analyses suggest integrating in-situ measurements and those provided by NASA Earth observations can be utilized in a multi-faceted SM evaluation, a valuable contribution to drought monitoring and water resource decision making.

Emma P Myrick↗
NASA NTRSDate not supplied

Tribology Induced Water Pump Bearing Failure

NASA’s In-Space Propulsion facility experienced two infant mortality water pump bearing failures during a recent refurbishment campaign, the cause or causes of which were unclear. A forensic failure study was undertaken that included site visits, collection and examination of physical evidence and a bearing and pump design review. The pumps are large (2000 hp) vertical turbine design machines that utilize metallic sleeve bearings cooled and lubricated by the pumped fluid (water). Though initial possible root cause for the failures were attributed to improper fabrication and debris contamination, detailed examination of failure surfaces combined with bearing design revealed that the failure mode was thermal seizure brought on by inadequate cooling flow. An unusual contributing factor was that the use of grease as an assembly lubricant, as opposed to a fluid like oil, appears to have exacerbated the rather than alleviated the thermal runaway by blocking water flow. Resolution of the problem was achieved through the design and installation of dedicated bearing lubrication water feed system.

bearings↗
DOE OSTI2024

Horizontal gene transfer of the Mer operon is associated with large effects on the transcriptome and increased tolerance to mercury in nitrogen-fixing bacteria

Abstract Background Mercury (Hg) is highly toxic and has the potential to cause severe health problems for humans and foraging animals when transported into edible plant parts. Soil rhizobia that form symbiosis with legumes may possess mechanisms to prevent heavy metal translocation from roots to shoots in plants by exporting metals from nodules or compartmentalizing metal ions inside nodules. Horizontal gene transfer has potential to confer immediate de novo adaptations to stress. We used comparative genomics of high quality de novo assemblies to identify structural differences in the genomes of nitrogen-fixing rhizobia that were isolated from a mercury (Hg) mine site that show high variation in their tolerance to Hg. Results Our analyses identified multiple structurally conserved merA homologs in the genomes of Sinorhizobium medicae and Rhizobium leguminosarum but only the strains that possessed a Mer operon exhibited 10-fold increased tolerance to Hg. RNAseq analysis revealed nearly all genes in the Mer operon were significantly up-regulated in response to Hg stress in free-living conditions and in nodules. In both free-living and nodule environments, we found the Hg-tolerant strains with a Mer operon exhibited the fewest number of differentially expressed genes (DEGs) in the genome, indicating a rapid and efficient detoxification of Hg from the cells that reduced general stress responses to the Hg-treatment. Expression changes in S. medicae while in bacteroids showed that both rhizobia strain and host-plant tolerance affected the number of DEGs. Aside from Mer operon genes, nif genes which are involved in nitrogenase activity in S. medicae showed significant up-regulation in the most Hg-tolerant strain while inside the most Hg-accumulating host-plant. Transfer of a plasmid containing the Mer operon from the most tolerant strain to low-tolerant strains resulted in an immediate increase in Hg tolerance, indicating that the Mer operon is able to confer hyper tolerance to Hg. Conclusions Mer operons have not been previously reported in nitrogen-fixing rhizobia. This study demonstrates a pivotal role of the Mer operon in effective mercury detoxification and hypertolerance in nitrogen-fixing rhizobia. This finding has major implications not only for soil bioremediation, but also host plants growing in mercury contaminated soils.

59 BASIC BIOLOGICAL SCIENCES↗