Biological Research In Canisters - 16: The Impact of Spaceflight on Arabidopsis: Deep Sequencing and DNA Arrays as Collaborative Readouts of the Transcriptome of Arabidopsis Seedlings and Undifferentiated Cells in Space (BRIC-16-DNA) - 12.17.14

Overview | Description | Applications | Operations | Results | Publications | Imagery
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Science Objectives for Everyone
Biological Research In Canisters - 16: The impact of spaceflight on Arabidopsis: Deep sequencing and DNA Arrays as Collaborative Readouts of the Transcriptome of Arabidopsis Seedlings and Undifferentiated Cells in Space (BRIC-16-DNA) compares and contrasts the gene expression responses within two forms of Arabidopsis: whole, etiolated seedlings and undifferentiated cells in culture. The comparison of intact plants with cultures of undifferentiated cells shows that cells can detect space flight and gravity in the absence of tissue or organized developmental structures.

Science Results for Everyone

Scientists know that plants adapt their behaviors in response to space flight, although previous studies on this have been limited. This investigation showed that seedlings and cell cultures in microgravity both alter specific gene expressions, but in different ways. The differences could be because seedlings use their organs to sense and sample their environment, while undifferentiated cell cultures lack such organs and so respond inappropriately to stress. Or, cells in the culture could be exhibiting identical responses, while distinct tissue-types in the seedlings each responded differently. Either way, further investigation is needed.



The following content was provided by Anna-Lisa Paul, Ph.D., and is maintained in a database by the ISS Program Science Office.

Experiment Details

OpNom BRIC

Principal Investigator(s)

  • Anna-Lisa Paul, Ph.D., University of Florida, Gainesville, FL, United States

  • Co-Investigator(s)/Collaborator(s)
  • Robert J. Ferl, Ph.D., University of Florida, Gainesville, FL, United States

  • Developer(s)
    Bionetics Corporation, Cape Canaveral, FL, United States

    Sponsoring Space Agency
    National Aeronautics and Space Administration (NASA)

    Sponsoring Organization
    Human Exploration and Operations Mission Directorate (HEOMD)

    Research Benefits
    Information Pending

    ISS Expedition Duration
    March 2010 - September 2010

    Expeditions Assigned
    23/24

    Previous ISS Missions
    ISS Expedition 23/24 is the first mission for BRIC-16-Regulation.

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    Experiment Description

    Research Overview

    • Biological Research In Canisters - 16: The Impact of Spaceflight on Arabidopsis: Deep sequencing and DNA Arrays as Collaborative Readouts of the Transcriptome of Arabidopsis Seedlings and Undifferentiated Cells in Space (BRIC-16-DNA) focuses on Arabidopsis, which is the preferred model species for plant investigations in space. Arabidopsis is a small plant with a short generation time and the advantages of a small genome size, a wealth of available genetic mutants, and an already well characterized pattern of development.


    • BRIC-16 flies Arabidopsis seeds and callus cultures on a Space Shuttle mission during which the seeds will germinate and the cultures grow and are subsequently returned to Earth when the Space Shuttle lands. Crew members will perform an in-flight chemical fixation to preserve the seedlings and cell cultures on-orbit prior to return.


    • These investigations will study how plants perceive and respond to gravity, and how gene regulation is altered by spaceflight conditions. The fundamental knowledge gained through these investigations will aid in our ability to better control plant use on Earth in agriculture (and other) applications.

    Description
    Providing a continuous supply of food, oxygen, and clean water for humans in space is a costly proposition. To date these needs have been met largely through stowage and resupply. As the durations of missions increase, the costs associated with this approach become prohibitive. For this reason, virtually all scenarios for long-term space missions involve plants as key components of the life support environment. Plants will be used to recycle wastes, remove carbon dioxide, purify water and produce oxygen and food for astronauts. Biological Research In Canisters - 16: The impact of spaceflight on Arabidopsis: Deep sequencing and DNA Arrays as Collaborative Readouts of the Transcriptome of Arabidopsis Seedlings and Undifferentiated Cells in Space (BRIC-16-DNA) compares and contrasts the gene expression responses within two forms of Arabidopsis: whole, etiolated seedlings and undifferentiated cells in culture. Two complementary types of genome-wide gene expression profiling analyses will be applied to the biological samples from both space flight and ground controls:

    • Replicated DNA microarray analyses that will reveal changes in genome-wide patterns of gene expression on a platforms whose data are directly comparable to the wide range of array data available for Arabidopsis.

    • RNA deep sequencing, RNASeq, a relatively new, digital transcript profiling method that offers a more accurate genome-wide quantitation of transcript abundance over wider dynamic range of expression levels.
    The baseline transcriptome profiling of seedlings and cultured cells addresses the fundamental hypothesis that space flight is a unique and challenging environment in which survival requires adaptive changes that are both governed and displayed by alterations in gene expression. The comparison of intact plants with cultures of undifferentiated cells addresses whether cells can detect space flight and gravity in the absence of tissue or organized developmental structures. These experiments will enable the evaluation of fundamental mechanisms associated with plant responses to the novel environment of space flight and answere the following questions:
    • What gene response pathways are engaged in response to space flight?

    • Are specific organs and tissues required for the detection and transduction of signals from the space flight environment, including gravity?

    • Given that certain differentiated cells are often associated with the perception or signaling of environmental events, what happens when they are absent?

    • Would gravity still exert an effect on plant cells?

    • Can single cells perceive gravity?
    These are key questions that underpin some of the biggest questions in space flight plant biology.

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    Applications

    Space Applications
    The fundamental knowledge gained by growing plants under microgravity conditions can contribute to resolving the following risks:

  • Providing and Maintaing Biodegenerative Life Support Systems

  • Maintaining Food Quantity and Quality

  • Maintaining Acceptable Atmosphere

  • Managing Waste

  • Providing and Recovering Potable Water
  • Earth Applications
    The microgravity of space will be used to investigate and clarify plant-related phenomena that cannot be studied in the presence of gravity. The fundamental knowledge gained through these investigations will aid in our understanding of basic plant processes that can eventually increase our ability to better control plant use on Earth in agriculture (and other) applications.

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    Operations

    Operational Requirements
    This is a passive payload, with no on-orbit power or communications available. The investigators will plate their biology onto 60 mm petri dishes containing agar-solidified media. Each petri dish will be placed inside its own Petri Dish Fixation Unit (PDFU). The PDFUs will be assembled and loaded with a fluid in the syringe department (as specified by the selected investigators). Five PDFUs plus one temperature data logger will be loaded into each BRIC-PDFU (Biological Research In Canisters - Petri Dish Fixation Unit). Preflight turnover will be no earlier than 24 hours prior to launch. In the event of a launch scrub, the entire assembly will be replaced with an identical back-up unit with freshly loaded biology. Crewmembers will perform one in-flight operation per petri dish (using actuator equipment) to chemically fix the tissues on-orbit prior to return using RNAlater. The PDFUs will remain contained within the BRIC-PDFUs during all phases of flight operations. The fixed samples will be subsequently returned to Earth for postflight processing.

    Operational Protocols
    Eight BRIC-PDFU canisters reside in ½ middeck locker drawer. During Actuation operations, the drawer is removed, and the BRIC canisters removed one at a time. The Actuator Tool and Rod Kit are removed from another middeck locker location. A single rod from the rod kit is inserted into the Actuator Tool. The Actuator Tool is used to insert the rod into a single location on a BRIC canister lid. Squeezing the handle on the Actuator Tool forces the rod through septa on the BRIC canister lid, then through septa on one of the PDFUs inside the canister. The rod forces a piston inside the PDFU to move fixatives from a storage volume into the Petri dish volume inside the PDFU. After Actuation, the rod remains contained inside the BRIC canister. The crew do not come in contact with fixatives, since the fixatives remain contained within 3 levels of containment throughout all phases of flight. The Actutation is repeated for all 5 PDFU locations within each of the eight BRICs, totaling 40 PDFUs, and 40 Actutations. The Actuation procedure is performed in 30 minutes. For the remainder of flight, the BRIC operates autonomously with no interface.

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    Results/More Information

    Previous studies determined that plants exhibit adaptive behaviors in response to space flight. These studies have been limited due to the lack of manifest opportunities and biological replication. BRIC-16-DNA addressed these issues by producing replicable results in order to answer fundamental questions using current gene profiling technologies. Seedlings and cell cultures both respond to microgravity by altering specific gene expressions in entirely different responses. A possible explanation for the differences between cultured cells and seedlings is due to the fact that seedlings use their organs to sense and sample their environment, and that undifferentiated cell cultures, lacking such organs propagate inappropriate stress responses. Another possibility is that all the cells in the culture were responding unanimously, while distinct tissue-types in the seedlings responded differently. Either way, further investigations will need to be held in order to make a more definite conclusion (Paul et al 2012). The research group is currently preparing a follow-on experiment with Arabidopsis tissue culture cells in BRIC-17.

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    Results Publications

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    Ground Based Results Publications

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    ISS Patents

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    Related Publications

      Ferl RJ, Ferl RJ, Laughner B.  In vivo detection of regulatory factor binding sites of Arabidopsis thaliana Adh. Plant Molecular Biology. 1989; 12: 257266.

      Paul A, Paul A, Bamsey M, Berinstain A, Braham S, Neron P, Murdoch T, Graham T, Ferl RJ, Ferl RJ.  Deployment of a Prototype Plant GFP Imager at the Arthur Clarke Mars Greenhouse of the Haughton Mars Project. Sensors and Actuators B: Chemical. 2008; 8: 27622773.

      Paul A, Paul A, Ferl RJ, Ferl RJ.  Higher order chromatin structures in maize and Arabidopsis. The Plant Cell. 1998; 10(8): 1349-1359. DOI: 10.1105/tpc.10.8.1349. PMID: 9707534.

      Paul A, Paul A, Ferl RJ, Ferl RJ.  Permeabilized Arabidopsis protoplasts provide new insight into the chromatin structure of plant alcohol dehydrogenase genes. Developmental Genetics. 1998; 22: 716.

      Paul A, Paul A, Liu L, McClung S, Laughner B, Chen S, Ferl RJ, Ferl RJ.  Comparative interactomics:analysis of arabidopsis 14-3-3 complexes reveals highly conserved 14-3-3 interactions between humans and plants.. Journal of Proteome Research. 2009; 8: 191319.

      Paul A, Paul A, Levine HG, McLamb W, Norwood KL, Reed DW, Stutte GW, Stutte GW, Wells HW, Ferl RJ, Ferl RJ.  Plant molecular biology in the space station era: Utilization of KSC fixation tubes with RNAlater. Acta Astronautica. 2005; 56: 623-628.

      Ferl RJ, Ferl RJ, Wheeler RM, Levine HG, Paul A, Paul A.  Plants in space. Current Opinion in Plant Biology. 2002; 5: 258263.

      Paul A, Paul A, Ferl RJ, Ferl RJ.  The Biology Of Low Atmospheric Pressure - Implications For Exploration Mission Design And Advanced Life Support. Gravitational and Space Biology. 2006; 19: 3-17.

      Paul A, Paul A, Daugherty CJ, Binh EA, Chapman DK, Norwood KL, Ferl RJ, Ferl RJ.  Transgene expression patterns indicate that spaceflight affects stress signal perception and transduction in arabidopsis. Plant Physiology. 2001; 126: 613621.

      Paul A, Paul A, Ferl RJ, Ferl RJ.  In vivo footprinting reveals unique cis-elements and different modes of hypoxic induction in maize Adh1 and Adh2. The Plant Cell. 1991; 3: 159168. DOI: 10.1105/tpc.3.2.159. PMID: 1840906.

      Paul A, Paul A, Schuerger AC, Popp MP, Richards JT, Manak MS, Ferl RJ, Ferl RJ.  Hypobaric biology: Arabidopsis gene expression at low atmospheric pressure. Plant Physiology. 2004; 134: 215223.

      Paul A, Paul A, Popp MP, Gurley WB, Guy C, Norwood KL, Ferl RJ, Ferl RJ.  Arabidopsis gene expression patterns are altered during spaceflight. Advances in Space Research. 2005; 36: 1175-1181.

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    Related Websites

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    Imagery

    image A. View of an assembled Petri Dish Fixation Unit (PDFU). B. Callus culture on petri dish within a PDFU prior to closure (courtesy of A-L Paul, as flown on STS-131).
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    image A. View of a PDFU on its side (launch position). B. Five PDFUs plus one temperature logger within a BRIC-PDFU canister prior to closure. C. BRIC-PDFU canister with two pin guards attached. D. Eight BRIC-PDFU canisters stowed within a half middeck locker (as flown on STS-131).
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    image Injection of fluids into PDFU petri dishes. A. Actuator Rod Kit and Actuator Tool. B. Actuator Tool attached to BRIC-PDFU canister after using the Actuator to push fluid into the PDFU.
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