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At least 235 records · Page 13
All sourcesNASA NTRSDOE OSTI
NASA NTRS1998

Tropospheric Wind Profile Measurements with a Direct Detection Doppler Lidar

Research has established the importance of global tropospheric wind measurements for large scale improvements in numerical weather prediction. In addition, global wind measurements provide data that are fundamental to the understanding and prediction of global climate change. These tasks are closely linked with the goals of the NASA Earth Science Enterprise and Global Climate Change programs. NASA Goddard has been actively involved in the development of direct detection Doppler lidar methods and technologies to meet the wind observing needs of the atmospheric science community. In this paper we describe a recently developed prototype wind lidar system using a direct detection Doppler technique for measuring wind profiles from the surface through the troposphere. This system uses a pulsed ND:YAG laser operating at 1064 nm as the transmitter. The laser pulse is directed to the atmosphere using a 40 cm diameter scan mirror. The portion of the laser energy backscattered from aerosols and molecules is collected by a 40 cm diameter telescope and coupled via fiber optics into the Doppler receiver. Single photon counting APD's are used to detect the atmospheric backscattered signal. The principle element of the receiver is a dual bandpass tunable Fabry Perot etalon which analyzes the Doppler shift of the incoming laser signal using the double edge technique. The double edge technique uses two high resolution optical filters having bandpasses offset relative to one another such that the 'edge' of the first filter's transmission function crosses that of the second at the half power point. The outgoing laser frequency is located approximately at the crossover point. Due to the opposite going slopes of the edges, a Doppler shift in the atmospheric backscattered laser frequency produces a positive change in signal for one filter and a negative change in the second filter. Taking the ratio of the two edge channel signals yields a result which is directly proportional to the component of the wind along the line-of-sight of the laser. Measuring the radial wind in several directions provides sufficient information to determine the true wind speed and direction. The lidar has operated from our laboratory at Goddard since June, 1997. Wind profiles have been obtained to altitudes of 12 km with a vertical resolution of 330 in. Vector wind data are obtained by rotating the scan mirror to measure line-of-sight wind profiles for at least two azimuth angles at an elevation angle of 45 degrees. The precision of the data as determined from the standard deviation of multiple independent lidar profiles is in the range of 1 to 3 m/sec up to 10 km. Good agreement is obtained when the lidar data are compared with the upper air rawinsonde soundings taken at Dulles airport. Examples of the wind lidar data will be presented along with a description of the instrument and future developments.

Gentry, Bruce M.↗
DOE OSTI2022

ARM TBS and TigerShark deployment data in 2021

In 2021, observational data collected using ARM platforms, including seven TigerShark UAS flights and 133 tethered balloon system (TBS) flights, were archived by the ARM Data Center and made publicly available to the user community.

54 ENVIRONMENTAL SCIENCES↗
NASA NTRS2018

Impact Study of the Assimilation of Surface Sensitive Microwave Radiances in the GEOS

The aim of this study is to investigate the feasibility of assimilating low frequency microwave observations from different satellite Microwave radiometers such as the Advanced Microwave Sounding Unit-A (AMSU-A). These observations are relevant to the description of air temperature, humidity, and surface parameters such as ocean surface temperature. Their assimilation into Goddard Earth Observing System (GEOS) modeling and assimilation system helps better constrain models in regions where very few observations are assimilated. In recent years, Channels 1–4 and 15 have not been assimilated in GEOS because of their large sensitivities to uncertain surface parameters such as emissivity and skin temperature. Here, the impact of using different channels will be discussed. Also we investigate the validity of the emissivity model in these channels over ocean and sea ice.Radiative transfer calculations necessary for the assimilation of satellite radiances are performed using the Community Radiative Transfer Model (CRTM). The CRTM performs forward radiative transfer calculations of brightness temperature from atmospheric profiles (temperature, moisture, and ozone) and surface characteristics. Additionally, the CRTM calculates the Jacobians of brightness temperature with respect to the input . The Jacobians are for the inversion of these radiances within the Gridpoint Statistical Interpolation (GSI) analysis system. Here, For direct assimilation of the radiance observations, the interface between GSI and CRTM, is slightly modified. Finally, we validate the assimilation results, and discuss the impact of each channels in detail.

Ebrahimi, Hamideh↗
DOE OSTI2021

Dissecting the dominant hot spring microbial populations based on community-wide sampling at single-cell genomic resolution

With advances in DNA sequencing and miniaturized molecular biology workflows, rapid and affordable sequencing of single-cell genomes has become a reality. Compared to 16S rRNA gene surveys and shotgun metagenomics, large-scale application of single-cell genomics to whole microbial communities provides an integrated snapshot of community composition and function, directly links mobile elements to their hosts, and enables analysis of population heterogeneity of the dominant community members. To that end, we sequenced nearly 500 single-cell genomes from a low diversity hot spring sediment sample from Dewar Creek, British Columbia, and compared this approach to 16S rRNA gene amplicon and shotgun metagenomics applied to the same sample. We found that the broad taxonomic profiles were similar across the three sequencing approaches, though several lineages were missing from the 16S rRNA gene amplicon dataset, likely the result of primer mismatches. At the functional level, we detected a large array of mobile genetic elements present in the single-cell genomes but absent from the corresponding same species metagenome-assembled genomes. Moreover, we performed a single-cell population genomic analysis of the three most abundant community members, revealing differences in population structure based on mutation and recombination profiles. While the average pairwise nucleotide identities were similar across the dominant species-level lineages, we observed differences in the extent of recombination between these dominant populations. Most intriguingly, the creek's Hydrogenobacter sp. population appeared to be so recombinogenic that it more closely resembled a sexual species than a clonally evolving microbe. Together, this work demonstrates that a randomized single-cell approach can be useful for the exploration of previously uncultivated microbes from community composition to population structure.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Contrasting Community Assembly Forces Drive Microbial Structural and Potential Functional Responses to Precipitation in an Incipient Soil System

Microbial communities in incipient soil systems serve as the only biotic force shaping landscape evolution. However, the underlying ecological forces shaping microbial community structure and function are inadequately understood. We used amplicon sequencing to determine microbial taxonomic assembly and metagenome sequencing to evaluate microbial functional assembly in incipient basaltic soil subjected to precipitation. Community composition was stratified with soil depth in the pre-precipitation samples, with surficial communities maintaining their distinct structure and diversity after precipitation, while the deeper soil samples appeared to become more uniform. The structural community assembly remained deterministic in pre- and post-precipitation periods, with homogenous selection being dominant. Metagenome analysis revealed that carbon and nitrogen functional potential was assembled stochastically. Sub-populations putatively involved in the nitrogen cycle and carbon fixation experienced counteracting assembly pressures at the deepest depths, suggesting the communities may functionally assemble to respond to short-term environmental fluctuations and impact the landscape-scale response to perturbations. We propose that contrasting assembly forces impact microbial structure and potential function in an incipient landscape; in situ landscape characteristics (here homogenous parent material) drive community structure assembly, while short-term environmental fluctuations (here precipitation) shape environmental variations that are random in the soil depth profile and drive stochastic sub-population functional dynamics.

16S amplicon sequencing↗
DOE OSTI2024

Microbial abundances and carbon use under ambient temperature or experimental warming in a southern boreal peatland

Abstract Organic peat soils occupy relatively little of the global land surface area but store vast amounts of soil carbon in northern latitudes where climate is warming at a rapid pace. Warming may result in strong positive feedbacks of carbon loss and global climate change driven by microbial processes if warming alters the balance between primary productivity and decomposition. To elucidate effects of warming on the microbial communities mediating peat carbon dynamics, we explored the abundance of broad microbial groups and their source of carbon (i.e. old carbon versus more recently fixed photosynthate) using microbial lipid analysis (δ 13 C PLFA) of peat samples under ambient temperatures and before/after initiation of experimental peat warming (+ 2.25, + 4.5, + 6.75, and + 9 °C). This analysis occurred over a profile to 2 m depth in an undrained, ombrotrophic peat bog in northern Minnesota. We found that the total microbial biomass and individual indicator lipid abundances were stratified by depth and strongly correlated to temperature under ambient conditions. However, under experimental warming, statistically significant effects of temperature on the microbial community were sporadic and inconsistent. For example, 3 months after experimental warming the relative abundance of Gram-negative bacterial indicators across depth combined and > 50 cm depth and Gram-positive bacterial indicators at 20–50 cm depth showed significant positive relationships to temperature. At that same timepoint, however, the relative abundance of Actinobacterial indicators across depth showed a significant negative relationship to temperature. After 10 months of experimental warming, the relative abundance of fungal biomarkers was positively related to temperature in all depths combined, and the absolute abundance of anaerobic bacteria declined with increasing temperature in the 20–50 cm depth interval. The lack of observed response in the broader microbial community may suggest that at least initially, microbial community structure with peat depth in these peatlands is driven more by bulk density and soil water content than temperature. Alternatively, the lack of broad microbial community response may simply represent a lag period, with more change to come in the future. The long-term trajectory of microbial response to warming in this ecosystem then could either be direct, after this initial lag time, or indirect through other physical or biogeochemical changes in the peat profile. These initial results provide an important baseline against which to measure long-term microbial community and carbon-cycling responses to warming and elevated CO 2 .

Felice, Mark↗
NASA NTRS2017

Exposing the Strategies that Can Reduce the Obstacles: Improving the Science User Experience

It is now well established that pursuing generic solutions to what seem are common problems in Earth science data access and use can often lead to disappointing results for both system developers and the intended users. This presentation focuses on real-world experience of managing a large and complex data system, NASAs Earth Science Data and Information Science System (EOSDIS), whose mission is to serve both broad user communities and those in smaller niche applications of Earth science data and services. In the talk, we focus on our experiences with known data user obstacles characterizing EOSDIS approaches, including various technological techniques, for engaging and bolstering, where possible, user experiences with EOSDIS. For improving how existing and prospective users discover and access NASA data from EOSDIS we introduce our cross-archive tool: Earthdata Search. This new search and order tool further empowers users to quickly access data sets using clever and intuitive features. The Worldview data visualization tool is also discussed highlighting how many users are now performing extensive data exploration without necessarily downloading data. Also, we explore our EOSDIS data discovery and access webinars, data recipes and short tutorials, targeted technical and data publications, user profiles and social media as additional tools and methods used for improving our outreach and communications to a diverse user community. These efforts have paid substantial dividends for our user communities by allowing us to target discipline specific community needs. The desired take-away from this presentation will be an improved understanding of how EOSDIS has approached, and in several instances achieved, removing or lowering the barriers to data access and use. As we look ahead to more complex Earth science missions, EOSDIS will continue to focus on our user communities, both broad and specialized, so that our overall data system can continue to serve the needs of science and applications users.

communications↗
NASA NTRSDate not supplied

Models, In situ, and Remote Sensing of Aerosols (MIRA)

There is a natural partitioning of scientific interest amongst three specialties of aerosol re-search: modeling, in situ measurements, and remote sensing. The broader aerosol community benefits when these groups interact, and this strengthens overall scientific impact on climate and air quality research and predictions. The new MIRA working group establishes a forum for identifying collaborations and improving discussions amongst specialties and across regional boundaries. One area of keen interest is uniting satellite and ground-based lidar groups with other aerosol disciplines. Elastic backscatter lidars depend upon a priori knowledge of aerosol properties to convert measured lidar profiles into aerosol extinction profiles. Acquiring additional insight on aerosol properties and transport is highly valuable to these lidar groups to improve the data quality and aid in their scientific interpretation. Another area of interest is to facilitate the incorporation of measurements into global aerosol models. Modelers need aerosol optical look-up tables that enable quick conversions of hydrated (and dry) polydisperse size distributions into aerosol optical properties (extinction, scattering, etc.), but many of the existing tables are based upon outdated measurements. Thus, MIRA is building the Tables of Aerosol Optics (TAO), which is a community collection of aerosol optical calculations. This expands on the historical efforts of Shettle and Fenn, d’Almeida, GADS, OPAC, etc., except that TAO seeks continual input from the community. Thus, as aerosol measurement and computational techniques advance, so does the TAO collective. Since TAO is a community collective, it is expected that users will optionally upload computations for aerosol type as well as computations for all of the traditional aerosol species(like amm sulfate, amm nitrate, organics, etc.). The near-term purpose of the MIRA working group is to: *Characterize regional aerosol lidar ratios to support improvements of aerosol extinction profiles and understanding of aerosol typing. *Create TAO --a community cooperative of aerosol optical tables. *Facilitate international communications between aerosol measurement and modeling groups. *Encourage the use of regional knowledge to develop and improve remote sensing techniques for current and future backscatter lidars located in space. *Enable and foster communication between the scientists who run global aerosol models and scientists who analyze space-based lidar data. We will discuss how a collaborative aerosol working group will be organized on this topic. Those who are interested in MIRA can sign up for the MIRA email listserver at https://forms.gle/qdbCnngzNJc5YJi57.

Greg Schuster↗
NASA NTRSDate not supplied

Models, In situ, and Remote sensing of Aerosols (MIRA): Formation of an International Working Group

There is a natural partitioning of scientific interest amongst three specialties of aerosol re- search: modeling, in situ measurements, and remote sensing. The broader aerosol community benefits when these groups interact, and this strengthens overall scientific impact on climate and air quality research and predictions. The new MIRA working group establishes a forum for identifying collaborations and improving discussions amongst specialties and across regional boundaries. One area of keen interest is uniting satellite and ground-based lidar groups with other aerosol disciplines. Elastic backscatter lidars depend upon a priori knowledge of aerosol properties to convert measured lidar profiles into aerosol extinction profiles. Acquiring additional insight on aerosol properties and transport is highly valuable to these lidar groups to improve the data quality and aid in their scientific interpretation. Another area of interest is to facilitate the incorporation of measurements into global aerosol models. Modelers need aerosol optical look-up tables that enable quick conversions of hydrated (and dry) polydisperse size distributions into aerosol optical properties (extinction, scattering, etc.), but many of the existing tables are based upon outdated measurements. Thus, MIRA is building the Tables of Aerosol Optics (TAO), which is a community collection of aerosol optical calculations. This expands on the historical efforts of Shettle and Fenn, d’Almeida, GADS, OPAC, etc., except that TAO seeks continual input from the community. Thus, as aerosol measurement and computational techniques advance, so does the TAO collective. Since TAO is a community collective, it is expected that users will optionally upload computations for aerosol type as well as computations for all of the traditional aerosol species (like amm sulfate, amm nitrate, organics, etc.). The near-term purpose of the MIRA working group is to: * Characterize regional aerosol lidar ratios to support improvements of aerosol extinction profiles and understanding of aerosol typing. * Create TAO -- a community cooperative of aerosol optical tables. * Facilitate international communications between aerosol measurement and modeling groups. * Encourage the use of regional knowledge to develop and improve remote sensing techniques for current and future backscatter lidars located in space. * Enable and foster communication between the scientists who run global aerosol models and scientists who analyze space-based lidar data. We will discuss how a collaborative aerosol working group will be organized on this topic. Send email to calipso_v5alr-join@lists.nasa.gov with the word `subscribe' in the subject line to join MIRA.

Gregory Schuster↗
DOE OSTI2021

A defined medium based on R2A for cultivation and exometabolite profiling of soil bacteria

SummaryExometabolomics is an approach to assess how microorganisms alter their environments through the depletion and secretion of chemical compounds. Comparisons of inoculated with uninoculated media can be used to provide direct biochemical observations on depleted and secreted metabolites which can be used to predict resource competition, cross-feeding and secondary metabolite production in microbial isolates and communities. This approach is most powerful when used with defined media that enable tracking of all depleted metabolites. However, microbial growth media have traditionally been developed for the isolation and growth of microorganisms but not metabolite utilization profiling through LC-MS/MS. Here, we describe the construction of a defined medium, the Northen Lab Defined Medium (NLDM), that not only supports the growth of diverse bacteria but is defined and therefore suited for exometabolomic experiments. Metabolites included in NLDM were selected based on their presence in R2A medium and soil, elemental stoichiometry requirements, as well as knowledge of metabolite usage by different bacteria. We found that NLDM supported the growth of 53 phylogenetically diverse soil bacterial isolates and all of its metabolites were trackable through LC–MS/MS analysis. These results demonstrate the viability and utility of the constructed NLDM medium for cultivating and characterizing diverse microbial isolates and communities.Originality-Significance StatementWe build a defined medium based on the metabolite composition of R2A medium and soil, elemental stoichiometry requirements, and knowledge of metabolite usage by different bacteria. The newly formulated defined medium was evaluated on its ability to support the growth of soil isolates and its application for metabolite utilization profiling. We found that of 53 phylogenetically diverse soil bacterial isolates grew on the defined medium and all of its metabolites were trackable through LC–MS/MS analysis. This demonstrates the viability and utility of the constructed defined medium for cultivating and characterizing diverse microbial isolates and communities.

de Raad, Markus↗
NASA NTRSDate not supplied

Large Scale Transcriptional Analysis of Legacy Spaceflight Tissues from the NASA Institutional Scientific Collection

The NASA Institutional Scientific Collection (ISC) has amassed a collection of valuable space biology samples spanning from early Space Shuttle missions to recent missions on the International Space Station (ISS). However, the full potential of this archive has not been realized, with many samples having been stored for decades without being re-accessed. Given the pace of analytical advancement since the ISC began accumulating samples, we undertook a systematic transcriptional analysis of these samples to reveal additional patterns that may have been missed during the original investigations. We selected 93 mouse and rat samples from 5 separate Space Shuttle, ISS and ground-analogue studies with a focus on muscle, spleen and thymus tissues to allow identification of important changes related to musculoskeletal unloading and immune function. RNA extracted from these tissues was consistently of a quality and we are now generating transcriptional profiling data from sample set. This resulting data will be immediately released through the GeneLab data systems for open analysis by the space biology community. While this study will stand on its own, it will also serve as a model for future, comprehensive, analyses of the ISC.

Rodent↗
DOE OSTI2025

In vitro activity and in vivo efficacy of omadacycline against Plasmodium species

Abstract Background Doxycycline is currently the only tetracycline-class antibiotic recommended for malaria prophylaxis. Omadacycline, a semisynthetic aminomethylcycline approved for treatment of adults with community-acquired bacterial pneumonia and acute bacterial skin and skin structure infections, has a well-established safety profile. This study evaluated the in vitro activity of omadacycline againstPlasmodium falciparumandPlasmodium cynomolgiand its in vivo efficacy againstPlasmodium bergheiin experimental malaria models to assess its potential as an antimalarial drug. Methods Fluorescence-based assays were used to assess the in vitro blood and liver stage activity of omadacycline and doxycycline againstP. falciparumandP. cynomolgilaboratory clones. In vivo liver and early-stage blood stage efficacy were evaluated in a murine model ofP. bergheiinfection, utilizing in vivo imaging of luciferase-expressingP. berghei(ANKA strain) sporozoites in female albino C57Bl/6 mice. Parasitaemia was monitored by flow cytometry for up to 30 days post-infection. Results Omadacycline demonstrated comparable in vitro activity to doxycycline against both drug-sensitive and drug-resistantP. falciparumclones, while doxycycline showed reduced activity against two drug-resistant clones. Notably, omadacycline exhibited superior anti-schizont activity in theP. cynomolgiliver stage assay. In theP. bergheimurine model, omadacycline was efficacious in both liver and early blood stages compared to the untreated control group, and demonstrated improved survival compared to doxycycline. Conclusions Omadacycline demonstrated enhanced antimalarial efficacy over doxycycline in vitro in liver stage activity and in overcoming resistance in the blood stage, and in survival in an in vivo model ofP. bergheiinfection. These findings support further investigation of omadacycline as a potential candidate for malaria prophylaxis and treatment.

Infectious Diseases↗
NASA NTRS2018

Microwave Remote Sensing of Snow: Advances over Ice Sheet, Land, and Sea Ice

Satellite microwave radiometers have enabled us to observe the cryosphere and its changes. A variety of algorithms have been developed since the late 1970s. These convert observed microwave radiation into geophysical, glaciological properties relevant to study ice sheets’ snow accumulation and melt, terrestrial snow water equivalent and freeze/thaw state, and various sea ice cover characteristics like extent, concentration, and thickness. In spite of the continuous availability of satellite microwave radiometers for the past 40 years, potential for advancing our understanding of the relationship between microwave radiation and snow/ice properties still exists. Original empirical algorithms have matured and are becoming more physically based. This presentation offers insights into some advances made during the past 10 years in monitoring ice sheet, terrestrial snow, and sea ice. These advances made it possible to provide new, more reliable climate-related variables to the community for the satellite era using the typical 18-37 GHz frequency range (e.g., retrievals of grain size profiles in Antarctica, snow cover stratification, and therefore accumulation with applications to climate studies). Recent NASA instruments have recorded low microwave frequencies (at 1.4 GHz). These observations have a large penetration depth, they emanate from deep into the ice, where properties including temperature are very stable. Nonetheless, it has been found at both Dome C, Antarctica and Summit, Greenland that changes in surface snow properties significantly influence these observations. Compared to ice sheets, terrestrial snow and sea ice present higher spatial heterogeneities. Over land, presence of canopy and lakes, though with known locations, add ambiguities in the retrievals of snow properties. Over sea ice, ridges, leads, changes in salinity, and sea ice drift augment further the level of difficulty in obtaining robust geophysical properties. Assessing the quality of satellite retrievals, often requiring field activities, is a necessary step in designing the next generation of microwave algorithms to monitor changes in the cryosphere.

Brucker, Ludovic↗
DOE OSTI2024

A functional microbiome catalogue crowdsourced from North American rivers

Predicting elemental cycles and maintaining water quality under increasing anthropogenic influence requires knowledge of the spatial drivers of river microbiomes. However, understanding of the core microbial processes governing river biogeochemistry is hindered by a lack of genome-resolved functional insights and sampling across multiple rivers. Here we used a community science effort to accelerate the sampling, sequencing and genome-resolved analyses of river microbiomes to create the Genome Resolved Open Watersheds database (GROWdb). GROWdb profiles the identity, distribution, function and expression of microbial genomes across river surface waters covering 90% of United States watersheds. Specifically, GROWdb encompasses microbial lineages from 27 phyla, including novel members from 10 families and 128 genera, and defines the core river microbiome at the genome level. GROWdb analyses coupled to extensive geospatial information reveals local and regional drivers of microbial community structuring, while also presenting foundational hypotheses about ecosystem function. Building on the previously conceived River Continuum Concept, we layer on microbial functional trait expression, which suggests that the structure and function of river microbiomes is predictable. We make GROWdb available through various collaborative cyberinfrastructures, so that it can be widely accessed across disciplines for watershed predictive modelling and microbiome-based management practices.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Next-Generation Intensity-Duration-Frequency Curves for Climate-Resilient Infrastructure Design: Advances and Opportunities

National and international security communities (e.g., U.S. Department of Defense) have shown increasing attention for innovating critical infrastructure and installations due to recurring high-profile flooding events in recent years. The standard infrastructure design approach relies on local precipitation-based intensity-duration-frequency (PREC-IDF) curves that do not account for snow process and assume stationary climate, leading to high failure risk and increased maintenance costs. This paper reviews the recently developed next-generation IDF (NG-IDF) curves that explicitly account for the mechanisms of extreme water available for runoff including rainfall, snowmelt, and rain-on-snow under nonstationary climate. The NG-IDF curve is an enhancement to the PREC-IDF curve and provides a consistent design approach across rain- to snow-dominated regions, which can benefit engineers and planners responsible for designing climate-resilient facilities, federal emergency agencies responsible for the flood insurance program, and local jurisdictions responsible for developing design manuals and approving subsequent infrastructure designs. Further, we discuss the recent advances in climate and hydrologic science communities that have not been translated into actional information in the engineering community. To bridge the gap, we advocate that building climate-resilient infrastructure goes beyond the traditional local design scale where engineers rely on recipe-based methods only; the future hydrologic design is a multi-scale problem and requires closer collaboration between climate scientists, hydrologists, and civil engineers.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2022

Habitat‐adapted microbial communities mediate Sphagnum peatmoss resilience to warming

Summary Sphagnum peatmosses are fundamental members of peatland ecosystems, where they contribute to the uptake and long‐term storage of atmospheric carbon. Warming threatens Sphagnum mosses and is known to alter the composition of their associated microbiome. Here, we use a microbiome transfer approach to test if microbiome thermal origin influences host plant thermotolerance. We leveraged an experimental whole‐ecosystem warming study to collect field‐grown Sphagnum , mechanically separate the associated microbiome and then transfer onto germ‐free laboratory Sphagnum for temperature experiments. Host and microbiome dynamics were assessed with growth analysis, Chl a fluorescence imaging, metagenomics, metatranscriptomics and 16S rDNA profiling. Microbiomes originating from warming field conditions imparted enhanced thermotolerance and growth recovery at elevated temperatures. Metagenome and metatranscriptome analyses revealed that warming altered microbial community structure in a manner that induced the plant heat shock response, especially the HSP70 family and jasmonic acid production. The heat shock response was induced even without warming treatment in the laboratory, suggesting that the warm‐microbiome isolated from the field provided the host plant with thermal preconditioning. Our results demonstrate that microbes, which respond rapidly to temperature alterations, can play key roles in host plant growth response to rapidly changing environments.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Facile One-Pot Nanoproteomics for Label-Free Proteome Profiling of 50–1000 Mammalian Cells

Recent advances in sample preparation enable label-free MS-based proteome profiling of small numbers of mammalian cells. However, specific devices are often required to downscale sample processing volume from the standard 50-200 µL to sub-µL for effective nanoproteomics, which greatly impedes the implementation of current nanoproteomics methods by broad proteomics research community. Here we report a facile one-pot nanoproteomics method termed SOPs-MS (Surfactant-assisted One-Pot sample processing at the standard volume coupled with MS) for convenient proteome profiling of 50-1000 mammalian cells. Building upon our recent development of SOP-MS for label-free single-cell proteomics at low µL volume (Commun Bio 2021, 4, 265), we have systematically evaluated its processing volume at 10-200 µL using 100 human cells for robust reproducible nanoproteomic analysis. The processing volume of 50 µL which is in the range of volume for standard proteomics sample preparation, has been selected for easy sample handling with benchtop micropipette. Using the commonly accessible LC-MS platform, SOPs-MS allows for reliable label-free quantification of ~1200-2700 protein groups from 50-1000 MCF10A cells. When applied to small subpopulations of mouse colon crypt cells, SOPs-MS can reveal distinct protein signatures between any two subpopulation cells with identification of ~1500-2500 protein groups for each subpopulation. SOPs-MS may pave the way for routine deep proteome profiling of small numbers of cells as well as low-input samples.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS2021

Characterization of Spacesuit Associated Microbial Communities and their Implications for NASA Missions

Crewed National Aeronautics and Space Administration (NASA) missions to other solar system bodies are currently being planned. One high-profile scientific focus during such expeditions would be life detection, specifically the discovery of past or present microbial life, if they exist. However, both humans and associated objects typically carry a high microbial burden. Thus, it is essential to distinguish between microbes brought with the expedition and those present on the exploring planets. Modern spacesuits are unique, customized spacecraft which provide protection, mobility and life support to crew during spacewalks, yet they vent continuously, and the mobility of microbes through spacesuits has not been studied. To evaluate the microbial colonization of spacesuits, NASA used an Extravehicular Activity swab kit to examine viable microbial populations of 48 samples from spacesuits using both traditional microbiological methods and molecular sequencing methods. The cultivable microbial population ranged from below the detection limit to 9 x 102 colony forming units per 25 cm2 of sample and also significantly varied by the location. The cultivable microbial diversity was dominated by members of Bacillus, Arthrobacter, and Ascomycota. However, 16S rRNA-based viable bacterial burden ranged from 105 to 106 copies per 25 cm2 of sample. Shotgun metagenome sequencing revealed the presence of a diverse microbial population on the spacesuit surfaces, including Curtobacterium and Methylobacterium from across all sets of spacesuits in high abundance. Among bacterial species identified, higher abundance of Cutibacterium acnes, Methylobacterium oryzae, and M. phyllosphaerae reads were documented. The results of this study provide evidence that identical microbial strains may live on the wrist joint, inner gauntlet, and outer gauntlet of spacesuits. This raises the possibility, but does not confirm, that viable microbes are able to migrate between the interior and exterior of spacesuits. Overall, these data provide the first estimate of microbial dispersion from internal to external spacesuit surfaces, which will help future mission planners develop effective planetary protection strategies.

spacesuit↗