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RNAseq analysis of the response of Arabidopsis thaliana phytochrome mutants (PhyA PhyB) to fractional gravity under blue-light stimulation during spaceflight
Traveling to nearby extraterrestrial objects having a reduced gravity level (partial gravity) compared to Earth s gravity is becoming a realistic objective for space agencies. The use of plants as part of life support systems will require a better understanding of the interactions among plant growth responses including tropisms under partial gravity conditions. Here we present results from the Seedling Growth space experiments on the ISS to complement the previously released GLDS-251 dataset including seeds of Arabidopsis thaliana wildtype plants. Seeds were germinated and seedlings grew for six days under different gravity levels namely micro-g several intermediate partial-g levels and 1g and were subjected to irradiation with blue light for the last 48 hours. RNA was extracted was obtained for 20 wildtype samples for subsequent RNAseq analysis in GLDS-251 here we add 36 samples from similarly exposed PhyA and PhyB mutants.
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The effect of spaceflight on transgenic Arabidopsis plants with compromised signaling
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Understanding the molecular mechanisms by which plants sense and adapt to changes in the space environment is essential for generating plants that are better adapted to withstand space flight microgravity and other adverse conditions encountered in space. The objective of our spaceflight experiment x93Plant Signaling in Microgravity x94 (carried out on the International Space Station ISS) was to compare transcript profiles of wild type and transgenic InsP 5-ptase plants with compromised InsP3 signaling. The transgenic Arabidopsis plants constitutively express the mammalian type I inositol polyphosphate 5-phosphatase (InsP 5-ptase) an enzyme that specifically hydrolyzes the lipid-derived second messenger inositol 1,4,5-trisphosphate (InsP3). These transgenic plants exhibit normal growth and morphology; however their responses to environmental stimuli including gravity and drought are altered. Seedlings were grown for 5 days under continuous light in experimental containers placed in the European Modular Cultivation system (EMCS) onboard the ISS. The EMCS consists of two rotors within a controlled chamber allowing for a x931g x94 control in space. After sample retrieval from the ISS RNA was isolated from shoot and root tissue and subjected to RNA sequencing. Two-way comparisons of micro g versus x931 x94g have uncovered regulatory mechanisms that are both conserved and altered between the wild type and transgenic seedlings.
Transcriptomics analysis of etiolated Arabidopsis thaliana seedlings in response to microgravity
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Gene expression profile of two-week-old etiolated Arabidopsis seedlings under microgravity on board space flight BRIC16 were compared with ground grown control in both wild-type and act2-3 mutant plants.
Comparison of the spaceflight transcriptome of four commonly used Arabidopsis thaliana ecotypes
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This experiment compared the spaceflight transcriptomes of four commonly used natural variants (ecotypes) of Arabidopsis thaliana using RNAseq. In nature Arabidopsis is a native of Europe/Asia/Northwestern Africa and is found across the globe growing in a wide range of environments. The geographical spread of these various populations has led to a slow divergence leading to distinct ecotypes. Understanding the impact of this ecotypic variability is an important factor when using Arabidopsis as a model. Seeds of the ecotypes Col_0 Ler-2 Ws-2 and Cvi-0 were flown to the International Space Station as part of CRS-4 mission in the Biological Research in Canister (BRIC) hardware. The seeds were germinated on orbit grown for 8 days and then fixed in RNAlater and frozen in the MELFI freezer for return to Earth. Once returned RNA was isolated and RNAseq performed to catalog the transcriptional patterns of the plants grown in space. An identical set of samples were grown in parallel on the ground to provide controls to allow assessment of transcriptional changes specifically associated with the spaceflight environment. This data release includes 48 out of 56 sample expression files with the remaining 8 files to be released at a later date.
Transcriptomic response of Drosophila melanogaster pupae developed in hypergravity
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Physical forces greatly influence the growth and function of an organism. Altered gravity can perturb normal development and induce corresponding changes in gene expression. Understanding this relationship between the physical and biological realms is important for NASA s space travel goals. We use combined RNA-Seq and qRT-PCR to profile changes in early Drosophila melanogaster pupae exposed to chronic hypergravity (3 g three times Earth s gravity) to highlight gravity-dependent pathways and gene products. Robust transcriptional response was evident among the pupae developed in a hypergravity environment compared to control. 1,513 genes showed significantly (p < 0.05) altered gene expression in the 3 g samples. These findings were supported with qRT-PCR data. Major biological processes affected include ion transport redox homeostasis immune and humoral stress response proteolysis and cuticle development.
Proteomics and Transcriptomics analysis of Arabidopsis Seedlings in Microgravity
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On Earth plants are constantly exposed to a gravitational field of 1g. Gravity affects a plant in every step of its development. Germinating seedlings orient their radicle and hypocotyl and growing plants position organs at a specific Gravitropic Set-point Angle dictated by the asymmetric distribution of auxin depending on the gravity vector. Hence gravitropism is one of the fundamental growth responses in plants. For any experiment studying the effects of gravity on plants the ultimate control is the microgravity in space. In this study Arabidopsis seeds were flown to the International Space Station and allowed to germinate and grow for 3 days in microgravity. Arabidopsis Wild Type Col-0 seeds were plated onto twenty-two 60mm Petri plates loaded into PDFUs and inserted 4 Biological Research in Canisters (BRICs). Approximately 800 seeds were sterilized plated on each 60mm Petri plates and cold stratified for 16 hours followed by 2 hours of white light treatment. The BRICs were maintained at 4C until spaceflight to ensure seed germination in microgravity. After 3 days of germination and growth the seedlings were fixed by injecting RNAlater into the chamber. They were kept at ambient temperature for 12 hours followed by freezing at -80C. An additional 22 plates were used as ground controls. After the spaceflight tissue from five plates was pooled to make each of three replicates. Both membrane and soluble proteins were extracted from the pooled seedlings. Proteins were trypsin digested labelled with iTRAQ and identified using tandem mass spectrometry.
Spaceflight adaptation requires organ specific alterations in the proteomes of Arabidopsis
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Life in spaceflight demonstrates remarkable adaptive processes within the specialized environments of space vehicles which are subject to the myriad of attending and unique environmental issues associated with orbital trajectories. To examine the adaptive processes that occur in plants in space leaves and roots from Arabidopsis seedlings that were grown from seed for 12 days on the International Space Station and preserved on orbit in RNAlater were returned to earth and analyzed using iTRAQ broad scale proteomics procedures.
Spaceflight adaptation requires organ specific alterations in the proteomes of Arabidopsis
공공데이터포털
Life in spaceflight demonstrates remarkable adaptive processes within the specialized environments of space vehicles which are subject to the myriad of attending and unique environmental issues associated with orbital trajectories. To examine the adaptive processes that occur in plants in space leaves and roots from Arabidopsis seedlings that were grown from seed for 12 days on the International Space Station and preserved on orbit in RNAlater were returned to earth and analyzed using iTRAQ broad scale proteomics procedures.
Transcriptomics analysis of etiolated Arabidopsis thaliana seedlings in response to microgravity
공공데이터포털
Gene expression profile of two-week-old etiolated Arabidopsis seedlings under microgravity on board space flight BRIC16 were compared with ground grown control in both wild-type and act2-3 mutant plants.
Biological Research in Canisters-16 (BRIC-16): Investigations of the plant cytoskeleton in microgravity with gene profiling and cytochemistry
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These investigations studied the fundamentals of how plants perceive gravity and develop in microgravity. It informs how gene regulation is altered by spaceflight conditions.
Candida albicans response to spaceflight (NASA STS-115)
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This study presents the first global transcriptional profiling and phenotypic characterization of the major human opportunistic fungal pathogen Candida albicans grown in spaceflight conditions. Microarray analysis revealed that C. albicans subjected to short-term spaceflight culture differentially regulated 454 genes compared to synchronous ground controls which represented 8.4% of the analyzed ORFs. Spaceflight-cultured C. albicans induced genes involved in cell aggregation (similar to flocculation) which was validated by microscopic and flow cytometry analysis. We also observed enhanced random budding of spaceflight-cultured cells as opposed to more normal bipolar budding patterns for ground samples in accordance with the gene expression data. Furthermore genes involved in antifungal agent and stress resistance were differentially regulated in spaceflight including induction of ABC transporters and members of the major facilitator family downregulation of ergosterol-encoding genes and upregulation of genes involved in oxidative stress resistance. Finally downregulation of genes involved in the actin cytoskeleton was observed. Interestingly the transcriptional regulator Cap1 and over 30% of the Cap1 regulon was differentially expressed in spaceflight-cultured C. albicans. A potential role for Cap1 in the spaceflight response of C. albicans is suggested as this regulator is involved in random budding cell aggregation actin cytoskeleton and oxidative stress resistance; all related to observed spaceflight-associated changes of C. albicans. While culture of C. albicans in microgravity potentiates a global change in gene expression that could induce a virulence-related phenotype no increased virulence in a murine intraperitoneal (i.p.) infection model was observed. This study represents an important basis for the assessment of the risk that commensal flora could play during spaceflight missions. Furthermore since the low fluid-shear environment of microgravity is relevant to physical forces encountered by pathogens during the infection process insights gained from this study could identify novel infectious disease mechanisms with downstream benefits for the general public. Cells were grown for 24 hours on the space shuttle or as ground-based controls preserved in RNALater and stored at -80C. Four samples of each flight- and ground-based controls were harvested for microarray analysis. GAP is Group Activation Pack and each GAP contains 8 FPAs. The numbers represent the # assigned to the particular GAP and the number assigned to the specific FPA (1-8) within the indicated GAP. The same hardware is used for the flight samples and the ground samples.