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SEEDLING GROWTH experiment was supported by the Spanish 'Plan Estatal de Investigaci\u00f3n Cient\u00edfica y T\u00e9cnica e Innovaci\u00f3n' of the Ministry of Economy, Industry and Competitiveness (Grant numbers AYA2012\u201333982 and ESP2015\u201364323-R to FJM, cofounded by ERDF). Funding for the RNA-seq was provided by NASA Grants NNX12A065G and 80NSSC17K0546 to JZK."},{"name":"Identifiers","value":""},{"name":"Managing NASA Center","value":"Ames Research Center (ARC)"},{"name":"Mission End","value":"03/26/2013, 10/25/2014"},{"name":"Mission Link","value":"https://osdr.nasa.gov/bio/repo/data/missions/SpaceX-2, https://osdr.nasa.gov/bio/repo/data/missions/SpaceX-4"},{"name":"Mission Name","value":"SpaceX-2, SpaceX-4"},{"name":"Mission Start","value":"03/01/2013, 09/21/2014"},{"name":"Project Identifier","value":"ARC00XX133"},{"name":"Project Link","value":"https://lsda.jsc.nasa.gov/Experiment/exper/1933"},{"name":"Project Title","value":"Novel Explorations Into the Interactions Between Light and Gravity Sensing in Plants - Seedling Growth-1"},{"name":"Project Type","value":"Spaceflight Study"},{"name":"Space Program","value":"NASA, ESA"},{"name":"Study Grant Number","value":""}],"description":"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 our latest space experiments on the ISS, in which seeds of Arabidopsis thaliana 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 from 20 samples for subsequent RNAseq analysis. Transcriptomic analysis was performed using the HISAT2-Stringtie-DESeq pipeline. Differentially expressed genes were further characterized for global responses using the GEDI tool, gene networks and for Gene Ontology (GO) enrichment.","factors":[{"@id":"rrAHixexWAdqOKNaj15Ezk66kWZSMcprcBRZfg1k","comments":[],"factorName":"Spaceflight","factorType":{"@id":"XJ2x9G5FYOhyGGbYDO0xkevuzcPxPnzdf1nFa9fv","annotationValue":"Space Flight","comments":[],"termAccession":"http://purl.bioontology.org/ontology/MESH/D013026","termSource":"MESH"}},{"@id":"XVwaJlTIF2q47sfrQq99a9PcHaoTfif10Dabpwg2","comments":[],"factorName":"Altered Gravity","factorType":{"@id":"PDmY1sKTXFdQ1hZVG6o5TrZ6BxH0kq1JMb1RuJZC","annotationValue":"Gravity, 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10-15 seedlings per extraction). A plant specific RNA extraction NucleoSpin kit (MACHEREY-NAGEL, Catalog # 740949.250) including a DNase treatment was used to isolate whole plant mRNA. The quantity and quality of the extracted RNA was determined by Nanodrop 2000 (Thermo Scientific). Extracted RNA was keep frozen at -80C until shipped on dry ice to the David H. Murdoch Research Institute in Kannapolis; North Carolina, USA.","name":"nucleic acid extraction","parameters":[{"@id":"#139742207913360","parameterName":{"@id":"Hop5C9AJDN9g3VYqtIJtB2A611oQjKFpzG6shAoZ","annotationValue":"QA Instrument","comments":[],"termAccession":"","termSource":""}}],"protocolType":{"@id":"zw8Tm4GuXt3NgUiREjJXSn2H3vXydRKOtV7U1XOV","annotationValue":"nucleic acid extraction","comments":[],"termAccession":"","termSource":""},"uri":"","version":""},{"@id":"#139742207911248","comments":[],"components":[],"description":"Twenty total RNA samples were used to generate twenty sequencing libraries using the Illumina TruSeq RNA Library Preparation Kit (Illumina, USA). 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Each pool was loaded onto a single lane of a flow cell. A 125bp paired end sequencing run was performed on the Illumina HiSeq2500.","name":"nucleic acid sequencing","parameters":[{"@id":"#139742268510112","parameterName":{"@id":"CbsOsBKhJANDso5kTtTkkEkONmwFxYPLJsPgkVT2","annotationValue":"sequencing instrument","comments":[],"termAccession":"","termSource":""}},{"@id":"#139742268507760","parameterName":{"@id":"Gpt4vDjDrU84g95ETwLXCbrKXjfl1JbGRu6ncg88","annotationValue":"Read Length","comments":[],"termAccession":"","termSource":""}}],"protocolType":{"@id":"4clR5NjwG34U2FF4v6tyx2puFqaEDOEYdcPGynJW","annotationValue":"nucleic acid sequencing","comments":[],"termAccession":"","termSource":""},"uri":"","version":""},{"@id":"#139742268508672","comments":[],"components":[],"description":"The raw fastq file for each sample was assessed for % rRNA using HTStream SeqScreener (version 1.0.0) and raw fastq file quality was evaluated with FastQC (version 0.12.1), and MultiQC (version 1.26) was used to generate raw MultiQC reports.","name":"GeneLab raw data processing protocol","parameters":[{"@id":"#139742268507712","parameterName":{"@id":"Ohs1BbENFJo4CWnwszKmpZ4g1nZJNqqJv60k0wyP","annotationValue":"Read Depth","comments":[],"termAccession":"http://ncicb.nci.nih.gov/xml/owl/EVS/Thesaurus.owl","termSource":"NCIT"}},{"@id":"#139742268508624","parameterName":{"@id":"RaWNd099ZqS9NO9TqfvpU9arANv1Do2iAJueQJMm","annotationValue":"rRNA Contamination","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}},{"@id":"#139742258114176","parameterName":{"@id":"XGdRyLdFBrQ5sINQ7N8WhowWQEznWY4WnzAmBL6l","annotationValue":"Merged Sequence Data","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}},{"@id":"#139742258110576","parameterName":{"@id":"WoqQnOe0wYp5oJdMtNlLbe7bcXgG6P1BgtOCncYd","annotationValue":"MultiQC File Names","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}}],"protocolType":{"@id":"gHRCpX8ZCZrvMb1jv0PpSihU4DurynDUUnkOSfet","annotationValue":"data transformation","comments":[],"termAccession":"http://purl.obolibrary.org/obo/OBI_0200000","termSource":"OBI"},"uri":"","version":""},{"@id":"#139742207911872","comments":[],"components":[],"description":"Data were processed as described in GL-DPPD-7101-G (<a href=\"https://github.com/nasa/GeneLab_Data_Processing/blob/master/RNAseq/Pipeline_GL-DPPD-7101_Versions/GL-DPPD-7101-G.md\" target=\"_\" >https://github.com/nasa/GeneLab_Data_Processing/blob/master/RNAseq/Pipeline_GL-DPPD-7101_Versions/GL-DPPD-7101-G.md</a>), using NF_RCP version 2.1.0 (https://github.com/nasa/GeneLab_RNASeq_Workflow/tree/NF_RCP_2.1.0). In short, raw fastq files were filtered using Trim Galore! (version 0.6.10) powered by Cutadapt (version 4.2). Trimmed fastq file quality was evaluated with FastQC (version 0.12.1), and MultiQC (version 1.26) was used to generate MultiQC reports. Arabidopsis Thaliana STAR and RSEM references were built using STAR (version 2.7.11b) and RSEM (version 1.3.3), respectively, Ensembl Plants release 59, genome assembly TAIR10 (Arabidopsis_thaliana.TAIR10.dna.toplevel.fa) and the following gtf annotation file: Arabidopsis_thaliana.TAIR10.59.gtf. Trimmed reads were aligned to the Arabidopsis Thaliana STAR reference with STAR (version 2.7.11b) and alignment log files were compiled with MultiQC (version 1.26). Aligned reads were assessed for strandedness using RSeQC infer_experiment.py (version 5.0.4), gene body coverage using RSeQC geneBody_coverage.py (version 5.0.4), inner distance was assessed using RSeQC inner_distance.py (version 5.0.4), and distribution of RNA features using RSeQC read_distribution.py (version 5.0.4). Aligned reads from all samples were quantified using RSEM (version 1.3.3), with strandedness set to none and RSEM count log files were compiled with MultiQC (version 1.26). To create a set of rRNA-removed quantification data, Ensembl IDs mapping to ribosomal RNA (rRNA) were removed from the RSEM gene count data. Both the original set of RSEM quantification data and the rRNA-removed data were subject to differential gene expression analysis as follows. A runsheet was generated with dp_tools (version 1.3.8) and the runsheet and quantification data were imported to R (version 4.4.2) with tximport (version 1.34.0) and normalized with DESeq2 (version 1.46.0) median of ratios method. Normalized gene counts were subject to differential expression analysis. Differential expression analysis was performed in R (version 4.4.2) using DESeq2 (version 1.46.0); all groups were compared pairwise using the Wald test and the likelihood ratio test was used to generate the F statistic p-value. Gene annotations were assigned using the custom annotation tables generated in-house as detailed in GL-DPPD-7110-A (<a href=\"https://github.com/nasa/GeneLab_Data_Processing/blob/GL_RefAnnotTable-A_1.1.0/GeneLab_Reference_Annotations/Pipeline_GL-DPPD-7110_Versions/GL-DPPD-7110-A/GL-DPPD-7110-A.md\" target=\"_\" >https://github.com/nasa/GeneLab_Data_Processing/blob/GL_RefAnnotTable-A_1.1.0/GeneLab_Reference_Annotations/Pipeline_GL-DPPD-7110_Versions/GL-DPPD-7110-A/GL-DPPD-7110-A.md</a>), with STRINGdb (version 2.16.4), PANTHER.db (version 1.0.12), and org.At.tair.db (version 3.19.1).","name":"GeneLab RNAseq data processing protocol","parameters":[{"@id":"#139742258113744","parameterName":{"@id":"3kGdO3NjDNgLooT0AsZzUCCBtnMbooOxqWyMLNIe","annotationValue":"trimmed sequence data","comments":[],"termAccession":"","termSource":""}},{"@id":"#139742258113312","parameterName":{"@id":"TTmlcsEgExQVnHKE7u07IoGATSJCxoPzxDzlwxpu","annotationValue":"aligned sequence data","comments":[],"termAccession":"","termSource":""}},{"@id":"#139742258114032","parameterName":{"@id":"Paz4Pjo6sV6E66RG3a9xuZDjKKpk7ywwq3VwmAwn","annotationValue":"Trimmed Sequence Data/MultiQC Reports","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}},{"@id":"#139742258113936","parameterName":{"@id":"s2HskUjUzSQmahcRYDfv1xBVjYScvewE8EO773hd","annotationValue":"Trimmed Sequence Data/Trimming Reports","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}},{"@id":"#139742258114080","parameterName":{"@id":"SPhWEr1D3r2VJ9OKO5b94MEY02Ipod6UwkLtQaRk","annotationValue":"Aligned Sequence Data/Unmapped Reads","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}},{"@id":"#139742250277952","parameterName":{"@id":"3AwnKZQDUPRaMntinM9bvBrGHpnb8qc6GaaZRIQh","annotationValue":"Aligned Sequence Data/Alignment Logs","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}},{"@id":"#139742250275936","parameterName":{"@id":"ib7AEGeVUoAruCpz91Qn6GXdO3E5d8k0DSeT2EQR","annotationValue":"Aligned Sequence Data/MultiQC Reports","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}},{"@id":"#139742250276128","parameterName":{"@id":"QcbiokEiY9OO2PO8IXfNhCwNUeMq4JM2ugyguLO3","annotationValue":"RSeQC/MultiQC Reports","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}},{"@id":"#139742250276992","parameterName":{"@id":"CzV3xNixKDCRnULWE97WbT9v456v77sgf4dPZG0Y","annotationValue":"Raw Counts Data","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}},{"@id":"#139742250275552","parameterName":{"@id":"WXhsQG8JLZ5oK8kUYbPxGSfXi520yz8A4dEHcBKs","annotationValue":"Raw Counts Tables","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}},{"@id":"#139742250278672","parameterName":{"@id":"w31o7Q91WYOVYIwRCqOCvdUs4H84WG2M5y8Qgu1N","annotationValue":"Raw Counts Data/MultiQC Reports","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}},{"@id":"#139742250277328","parameterName":{"@id":"Lns3jxVPjBQ2dUuFO7KyPzSqST1hCHcuYbzwld4i","annotationValue":"Normalized Counts Data","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}},{"@id":"#139742250276320","parameterName":{"@id":"c4UoiNar4sA4ty0VawH08rhaf4AR7F8drWBP91tt","annotationValue":"differential expression analysis data","comments":[],"termAccession":"http://purl.obolibrary.org/obo/OBI_0002584","termSource":"OBI"}}],"protocolType":{"@id":"gHRCpX8ZCZrvMb1jv0PpSihU4DurynDUUnkOSfet","annotationValue":"data transformation","comments":[],"termAccession":"http://purl.obolibrary.org/obo/OBI_0200000","termSource":"OBI"},"uri":"","version":""},{"@id":"#139742268508912","comments":[],"components":[],"description":"Seedlings were illuminated under white light (30-40 umol m-2 s-1) for 96 h, followed by 48 hours of unidirectional photostimulation with blue light. Light sources were LEDs. Spaceflight experiments were conducted utilizing the European Modular Cultivation System (EMCS) in the Columbus Module of the ISS.  The EMCS facility provides two centrifuges for creation of simulated gravity vectors, as well as atmospheric, temperature and hydration monitoring and control.","name":"growth protocol","parameters":[{"@id":"#139742257694896","parameterName":{"@id":"rPGDn3yIvLMOtVNtO0GUCMk2lds483aU4OXnZ7e1","annotationValue":"Hardware","comments":[],"termAccession":"https://genelab-data.ndc.nasa.gov/","termSource":""}},{"@id":"#139742257693264","parameterName":{"@id":"uRXa2oN701DjkFUq0vnYXDk8xa4w6ppyZhPDjkhG","annotationValue":"growth environment","comments":[],"termAccession":"http://purl.obolibrary.org/obo/OBI_0000997","termSource":"OBI"}},{"@id":"#139742257693072","parameterName":{"@id":"3eJ4QJs154TXdIdLiDlu5RGIZSFgVuhZDvPNF5eR","annotationValue":"light source","comments":[],"termAccession":"http://purl.obolibrary.org/obo/OBI_0400065","termSource":"OBI"}},{"@id":"#139742257693024","parameterName":{"@id":"e69rwVWyS2T1wImHQKexDAYuFTaDTE9k06hp0nVV","annotationValue":"Light Intensity","comments":[],"termAccession":"https://genelab-data.ndc.nasa.gov/","termSource":""}},{"@id":"#139742257693120","parameterName":{"@id":"KYfj5W8EUWSlT51HcJp7uPOjZ1ZBFefKS1y2EkvO","annotationValue":"Growth Medium","comments":[],"termAccession":"http://ncicb.nci.nih.gov/xml/owl/EVS/Thesaurus.owl","termSource":"NCIT"}},{"@id":"#139742257693408","parameterName":{"@id":"N7Xno1TWrsVfaXexERZmPlrty3SwUUid5sRUENlq","annotationValue":"Growth Light Color","comments":[],"termAccession":"https://genelab-data.ndc.nasa.gov/","termSource":""}},{"@id":"#139742257694848","parameterName":{"@id":"8oDvD75qlf6SmGKHsEgsZXmOJUkAEz1PePk1j2Dg","annotationValue":"Growth Time","comments":[],"termAccession":"https://genelab-data.ndc.nasa.gov/","termSource":""}},{"@id":"#139742257694560","parameterName":{"@id":"gPoyKxutsYx1S4iXDYb0Zot8s7u4NvDrrBfzkxz2","annotationValue":"Growth Time Start Date (MM/DD/YY)","comments":[],"termAccession":"https://genelab-data.ndc.nasa.gov/","termSource":""}}],"protocolType":{"@id":"0gMEkcMNeDUnXJOXyMiO3HWs7cSThiG9RpoYliDD","annotationValue":"growth protocol","comments":[],"termAccession":"http://www.ebi.ac.uk/efo/EFO_0003789","termSource":"EFO"},"uri":"","version":""},{"@id":"#139742268506464","comments":[],"components":[],"description":"Seeds of Arabidopsis thaliana ecotype Landsberg erecta (Ler) were flown to the ISS via the SpaceX Dragon. Spaceflight experiments were conducted utilizing the European Modular Cultivation System (EMCS) in the Columbus Module of the ISS.  The EMCS facility provides two centrifuges for creation of simulated gravity vectors, as well as atmospheric, temperature and hydration monitoring and control (Brinckmann and Schiller, 2002;Brinckmann, 2005;Kiss et al., 2014). In addition, the EMCS contains a video camera for image acquisition as well as monitoring of growth.  The Seedling Growth series of experiments was conducted in two parts. The first set of seedlings were uploaded on SpaceX CRS-2 (March 2013) followed by return via CRS-3 (May 2014), and the second set of seedlings were carried to the ISS on SpaceX CRS-4 (September 2014) and returned on CRS-5 (February 2015).  Experimental containers were uploaded to the ISS and loaded into the EMCS as previously described (Kiss et al., 2014;Vandenbrink and Kiss, 2016;Vandenbrink et al., 2019). Experimental conditions were controlled remotely from the Norwegian User Support and Operations Centre (N-USOC; Trondheim, Norway).  The experiment was initiated via hydration of the seeds.  Plants were grown under 6 nominal gravity conditions produced by different rotational speeds on the EMCS centrifuge, microgravity (stopped EMCS centrifuge), 0.1-g, 0.3-g, 0.5-g, 0.8-g and 1.0-g. The angular speed to generate each fractional gravity level was calculated for the cassette in the center of the Experimental Container. In the case of the 1.0-g cassettes, the value was calculated for the fifth cassette in order to prevent values higher than Earth nominal gravity.","name":"treatment protocol","parameters":[{"@id":"#139742257692928","parameterName":{"@id":"2AqoUNvqcX3D8TK1amME5Uxp5LIUN3PrpkTx5fPE","annotationValue":"Actual Gravity Level","comments":[],"termAccession":"","termSource":""}},{"@id":"#139742257694704","parameterName":{"@id":"XiKdsJvHTH9sIku341Brpb3Nu6C6k7E3Y186ypgW","annotationValue":"Light Treatment Duration","comments":[],"termAccession":"https://genelab-data.ndc.nasa.gov/","termSource":""}},{"@id":"#139742257693984","parameterName":{"@id":"E2itNn0MJVHa2Kpqy1sxVjIYBIsstuOwOuEuC1qG","annotationValue":"Treatment Light Color","comments":[],"termAccession":"https://genelab-data.ndc.nasa.gov/","termSource":""}},{"@id":"#139742257694752","parameterName":{"@id":"snTqVXJ3dYWlR2zyy8qyfvRX2sYmVJmK72MGnRI1","annotationValue":"Treatment Light Intensity","comments":[],"termAccession":"https://genelab-data.ndc.nasa.gov/","termSource":""}},{"@id":"#139742257695424","parameterName":{"@id":"EszLkJ7VvPWthZAwd1gmX67HusZTtaIMBrJJVv7u","annotationValue":"Treatment Light Source","comments":[],"termAccession":"https://genelab-data.ndc.nasa.gov/","termSource":""}},{"@id":"#139742257696624","parameterName":{"@id":"2SbNLUMxrYeqvAVo3Mt0aokRcjZKFpkrZcTRcU0A","annotationValue":"Gravity Treatment Duration","comments":[],"termAccession":"https://genelab-data.ndc.nasa.gov/","termSource":""}},{"@id":"#139742257693744","parameterName":{"@id":"xDdR5rcCNpWGPu2xr4lL7wekfuZo3YkGNcQ1QvMK","annotationValue":"Mission Launch","comments":[],"termAccession":"https://genelab-data.ndc.nasa.gov/","termSource":""}},{"@id":"#139742257695184","parameterName":{"@id":"9WDCCUkQla545cWMNT7Qe0CSPDSAiRSKXoPitIX4","annotationValue":"Mission Return","comments":[],"termAccession":"https://genelab-data.ndc.nasa.gov/","termSource":""}},{"@id":"#139742257693840","parameterName":{"@id":"i7ynUn4yQPoB59SFZkk4SY3WIEHSz6sOukWvTHII","annotationValue":"Launch Date","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":""}}],"protocolType":{"@id":"ZgJxEMQAhDfdpfObqMyljXQyrGlovo2dP3bZqLfq","annotationValue":"treatment protocol","comments":[],"termAccession":"","termSource":"EFO"},"uri":"","version":""},{"@id":"#139742258111200","comments":[],"components":[],"description":"The absorbed dose of ionizing radiation received by biological payloads on the International Space Station (ISS) depends on a number of factors, including date and duration of exposure and location of the payload on the ISS. Some of the factors that lead to variations in the absorbed dose rate in orbit are ISS altitude, period during the solar cycle, and location of the sample\u2014the ISS module and the specific location within that module.  Shielding effects from the biological payload container or habitat have not been taken into account.\n\nThe measured absorbed dose can vary based on the dosimetry instrument(s) used. Radiation monitoring on the ISS is not standardized across modules; each international partner has its own suite of instruments, with differing technologies, sensitivities and analysis methodologies, including criteria for distinguishing between lightly ionizing particles (mainly protons) and highly ionizing particles (mainly galactic cosmic ray nuclei).\n\nBoth active and passive dosimeters are used.  Active dosimeters are powered and data are read out and stored and/or telemetered to ground. Some active dosimeters provide additional levels of detail such as time resolution, energy deposition and particle identification.  Passive dosimeters are unpowered, record constantly and are read out upon return to the ground.\n\nThe dose in the metadata is the dose recorded by the radiation detector closest to the sample. In most cases that detector will be in the same ISS module; in cases where data from a detector in the same module are not available, the dose is extrapolated using data from periods where data are available from detectors in multiple modules, including the module containing the payload.\n\nFor plant studies the experiment duration begins at the time of onset of conditions that allow for seed germination. Once the specimen was frozen or fixed, the experiment was considered stopped.  Details of ISS dosimetry and a table with doses for all GeneLab ISS experiment ISS datasets are available at https://www.nasa.gov/reference/osdr-environmental-data/.\n\nThe first set of seedlings (SG1) were uploaded on SpaceX CRS-2 (March 2013) and returned on CRS-3 (May 2014), and the second set of seedlings (SG2)  were carried to the ISS on SpaceX CRS-4 (September 2014) and returned on CRS-5 (February 2015). Dates from R. Herranz (priv. comm.). The total mission duration of SG1 was ~443 days and SG2 was ~147 days. Seeds were dormant at launch, then germinated and grown for six days, then harvested and frozen. Absorbed radiation dose and dose rate reported are from the germination to harvest. 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