Microgravity effect on C. elegans gene expression was analysed by whole genome microarray. The worms were cultivated under microgravity for 4 days in the Japanese Module of the International Space Station. C. elegans N2 was exposed microgravity for 4 days. The worms synchronously were cultivated from L1 larvae to adult. There are two control groups onboard 1G and ground 1G control.

Space radiations and microgravity both could cause DNA damage in cells but the effects of microgravity on DNA damage response to space radiations are still controversial. A mRNA microarray and microRNA microarray in dauer larvae of Caenorhabditis elegans (C. elegans) that endured space xef xac x82ight environment and space radiations environment during 16.5-day Shenzhou-8 space mission were performed. In our study wild type and ced-1 mutant strains of C.elegans endured three conditions during shenzhou-8 spaceflight mission including spaceflight static condition(ss) spaceflight 1-g centrifugal condition(sc) and ground control condition(gc). Limited to the quantity of worm samples we performed technical-repeat test but not sample-repeat test.Accordingly xef xbc x8csix miRNA microarrays were performed.

Space radiations and microgravity both could cause DNA damage in cells but the effects of microgravity on DNA damage response to space radiations are still controversial. A mRNA microarray and microRNA microarray in dauer larvae of Caenorhabditis elegans (C. elegans) that endured space xef xac x82ight environment and space radiations environment during 16.5-day Shenzhou-8 space mission were performed. In our study wild type and dys-1 mutant strains of C.elegans endured four conditions during shenzhou-8 spaceflight mission including spaceflight static condition(ss) spaceflight 1-g centrifugal condition(sc) ground control condition(gc) and no-transport control. Limited to the quantity of worm samples we performed technical-repeat test but not sample-repeat test. Accordingly eight miRNA microarrays were performed.

Epigenetic changes in the DNA methylome are increasingly shown to play an integral role in regulating gene expression necessary for plants adaption to environmental stressors. Plants subjected to the novel environment of spaceflight onboard the International Space Station (ISS), show stress-related transcriptomic changes most notably associated with pathogen stress response. Here, we investigate how known terrestrial stress associated epigenetic modulations might play a role in spaceflight adaptation. To examine the role of 5mCyt in spaceflight adaptation, the APEX04-EPEX experiment conducted onboard the ISS evaluated the spaceflight altered genome wide methylation profiles of two methylation regulating gene mutants (methyltransferase 1 (met1-7) and elongator complex subunit 2 (elp2-5) that are involved in pathogen defense response, along with a wild type Col-0 control. MethylSeq and RNAseq analyses were performed on both spaceflight grown samples and ground grown controls. In addition, the epigenetics effects that may contribute to the differential gene expression patterns observed between leaf and root tissues were also investigated in an organ-specific manner.

The Advanced Plant Experiment-04 - Epigenetic Expression (APEX-04-EpEx) experiment onboard the International Space Station examined the spaceflight-altered cytosine methylation in two genetic lines of Arabidopsis thaliana, wild-type Col-0 and the mutant elp2-5, which is deficient in an epigenetic regulator Elongator Complex Subunit 2 (ELP2). Whole-genome bisulfite sequencing (WGBS) revealed distinct spaceflight associated methylation differences, presenting the need to explore specific space-altered methylation at single-molecule resolution to associate specific changes over large regions of spaceflight related genes. To date, tools of multiplexed targeted DNA methylation sequencing remain limited for plant genomes. This data set includes single-molecule profiling in user-defined targets using Flap-Enabled Next-Generation Capture (FENGC) on Arabidopsis root tissues to reveal precise modification of DNA methylation regulated by Elongator Complex Subunit 2 during spaceflight.

Epigenetic changes in the DNA methylome are increasingly shown to play an integral role in regulating gene expression necessary for plants’ adaption to environmental stressors. Plants subjected to the novel environment of spaceflight onboard the International Space Station (ISS), show stress-related transcriptomic changes most notably associated with pathogen stress response. Here, we investigate how known terrestrial stress associated epigenetic modulations might play a role in spaceflight adaptation. To examine the role of 5mCyt in spaceflight adaptation, the APEX04-EPEX experiment conducted onboard the ISS evaluated the spaceflight altered genome wide methylation profiles of two methylation regulating gene mutants, methyltransferase 1 (met1-7) and elongator complex subunit 2 (elp2-5), that are involved in pathogen defense response, along with a wild type Col-0 control. MethylSeq and RNAseq analyses were performed on both spaceflight grown samples and ground grown controls. In addition, the epigenetics effects that may contribute to the differential gene expression patterns observed between leaf and root tissues were also investigated in an organ-specific manner.

Experimentation on the International Space Station has reached the stage where repeated and nuanced transcriptome studies are beginning to illuminate the structural and metabolic differences between plants grown in space compared to plants on the Earth. Genes that are important in setting up the spaceflight responses are being identified; their role in spaceflight physiological adaptation are increasingly understood and the fact that different genotypes adapt differently is recognized. However the basic question of whether these spaceflight responses are required for survival has yet to be posed and the fundamental notion that spaceflight responses may be non-adaptive has yet to be explored. Therefore the experiments presented here were designed to ask if portions of the plant spaceflight response can be genetically removed without causing loss of spaceflight survival and without causing increased stress responses. The CARA experiment compared the spaceflight transcriptome responses of two Arabidopsis ecotypes Col-0 and WS as well as that of a PhyD mutant of Col-0. When grown with the ambient light of the ISS phyD displayed a significantly reduced spaceflight transcriptome response compared to Col-0 suggesting that altering the activity of a single gene can actually improve spaceflight adaptation by reducing the transcriptome cost of physiological adaptation. The WS genotype showed an even simpler spaceflight transcriptome response in the ambient light of the ISS more broadly indicating that the plant genotype can be manipulated to reduce the transcriptome cost of plant physiological adaptation to spaceflight and suggesting that genetic manipulation might further reduce or perhaps eliminate the metabolic cost of spaceflight adaptation. When plants were germinated and then left in the dark on the ISS the WS genotype actually mounted a larger transcriptome response than Col-0 suggesting that the in-space light environment affects physiological adaptation which further implies that manipulating the local habitat can also substantially impact the metabolic cost of spaceflight adaptation.

Expanding human spaceflight beyond low-Earth orbit poses significant technological challenges. One of these challenges is environmental monitoring, specifically the ability to identify and respond to microbiological threats in real-time. Because returning samples to Earth for analysis is not practical from deep-space, in-flight environmental monitoring is an essential capability for missions away from low-Earth orbit. Microbial identification by direct sequencing is readily achieved by identifying species-specific differences in Deoxyribonucleic Acid (DNA) sequences. These analyses require the extraction and separation of DNA from cells, but current methods for DNA isolation typically require technologies incompatible with spaceflight.

In-flight environmental monitoring is an important technology for missions away from low-Earth orbit. An important aspect of environmental monitoring is the ability to identify and respond to microbiological threats. Automatable in situ monitoring technologies are essential when sample return is not practical. Microbial identification is readily achieved by identifying differences in DNA sequences, such as direct sequencing or hybridization of the DNA strands. These analyses require the extraction and separation of DNA from cells. However, current methods for DNA isolation typically require technologies incompatible with spaceflight.

We seek to develop a miniaturized MEDIUM device for the extraction of DNA from operationally relevant samples (i.e., potable water sources); the extracted DNA can then be used for microbial identification using one or more DNA-dependent molecular techniques, such as sequencing. Isolation of DNA for sequencing or other analyses typically requires technologies incompatible with spaceflight. For this reason we have carefully chosen digital microfluidics-based extraction cartridges due to their likely compatibility with microgravity. Unlike other DNA extraction methods that rely on gravitational forces, the microfluidics devices use electro-wetting to move liquids. This eliminates the requirement for pumps, centrifuges and other moving parts. The quality of the DNA extracted by the microfluidics-based platform and by conventional kit-based methods will be characterized for its suitability in next-generation sequencing applications.

The goal of this study was to assess whether low shear-modeled microgravity (LSMMG) affects yeast genomic expression patterns using the powerful tool of whole genome microarray hybridization. We determined changes in the yeast model organism, Saccharomyces cerevisisae, when grown in LSMMG using the rotating High Aspect Ratio Vessel (HARV). A significant number of genes were up- or down-regulated by at least two fold in cells that were grown for 5 generations or 25 generations in HARVs. We identified genes in cell wall integrity signaling pathways containing MAP kinase cascades that may provide clues to novel physiological responses of eukaryotic cells to the external stress of a low-shear modeled microgravity environment. A comparison of the microgravity response to other environmental stress response (ESR) genes showed that 26% of the genes that respond ,significantly to LSMMG are involved in a general environmental stress response, while 74% of the genes may represent a unique transcriptional response to microgravity. In addition, we found changes in genes involved in budding, cell polarity establishment, and cell separation that confirm our hypothesis that exposure to LSMMG causes changes in gene transcription resulting in a phenotypic response. The results of the study provide interesting clues to potential mechanisms involved in the response to, adaptation to, and survival of eukaryotic cells in a microgravity environment and our findings may have important health implications for human spaceflight. Experiment Overall Design: Four conditions are compared with three replicates each: yeast grown in low-shear modeled microgravity (HARV bioreactor) for 5 and 25 generations; yeast grown in a horizontal (non-LSMMG) HARV bioreactor for 5 and 25 generations.

Premise of the study: The root apex is an important region involved in environmental sensing, but comprises a very small part of the root. Obtaining root apex transcriptomes is therefore challenging when the samples are limited. The feasibility of using tiny root sections for transcriptome analysis was examined, comparing RNA sequencing (RNA-Seq) to microarrays in characterizing genes that are relevant to spaceflight.Methods:Arabidopsis thaliana Columbia ecotype (Col-0) roots were sectioned into Zone 1 (0.5 mm; root cap and meristematic zone) and Zone 2 (1.5 mm; transition, elongation, and growth-terminating zone). Differential gene expression in each was compared.Results: Both microarrays and RNA-Seq proved applicable to the small samples. A total of 4180 genes were differentially expressed (with fold changes of 2 or greater) between Zone 1 and Zone 2. In addition, 771 unique genes and 19 novel transcriptionally active regions were identified by RNA-Seq that were not detected in microarrays. However, microarrays detected spaceflight-relevant genes that were missed in RNA-Seq. Discussion: Single root tip subsections can be used for transcriptome analysis using either RNA-Seq or microarrays. Both RNA-Seq and microarrays provided novel information. These data suggest that techniques for dealing with small, rare samples from spaceflight can be further enhanced, and that RNA-Seq may miss some spaceflight-relevant changes in gene expression.