Biological Research In Canisters - 16: Investigations of the plant cytoskeleton in microgravity with gene profiling and cytochemistry (BRIC-16-Cytoskeleton) - 08.24.16

Overview | Description | Applications | Operations | Results | Publications | Imagery

ISS Science for Everyone

Science Objectives for Everyone
Biological Research In Canisters - 16: Investigations of the plant cytoskeleton in microgravity with gene profiling and cytochemistry (BRIC-16-Cytoskeleton) studies the effects of microgravity on the structure and organization of the actin cytoskeleton in plants using the model plant Arabidopsis. The specific aims of this research are: to investigate plastid position in statocytes (the gravity-perceiving cells) in microgravity; to determine the effect of microgravity on the actin cytoskeletal organization in gravity-perceiving cells; to study microgravity effects on actin cytoskeleton-related gene expression in plant cells.
Science Results for Everyone
Seedlings grown in total darkness in microgravity showed more skewing at the roots and more roots forming from shoot tissues than ground control plants. But seeds in flight hardware in space and on the ground produced significantly fewer plants and had significantly smaller endodermal cells than those grown in petri dishes. This indicates alterations in cell wall that appear to be a true microgravity effect and suggests that the sealed canister system may have detrimental effects on seedling growth. Researchers therefore recommend caution in interpreting future results from the system and additional ground controls to separate space-hardware and true microgravity effects.

The following content was provided by John Z. Kiss, Ph.D., and is maintained in a database by the ISS Program Science Office.
Experiment Details

OpNom:

Principal Investigator(s)
John Z. Kiss, Ph.D., University of North Carolina-Greensboro, Greensboro, NC, United States

Co-Investigator(s)/Collaborator(s)
Richard E. Edelmann, Ph.D., Miami University, Oxford, OH, 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 Missions
ISS Expedition 23/24 is the first mission for BRIC-16-Cytoskeleton.

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Experiment Description

Research Overview

  • This payload focuses on Arabidopsis, 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 on a Space Shuttle mission during which the seeds will germinate and grow until returned to Earth on the Space Shuttle. Crewmembers will perform an in-flight chemical fixation to preserve the tissues 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 BRIC-16-Cytoskeleton studies the effects of microgravity on the structure and organization of the actin cytoskeleton in plants. This investigation builds on previous ground-based and space flight research using the model plant Arabidopsis. Thus, the specific aims of this proposed flight research are to investigate plastid position in statocytes (the gravity-perceiving cells) in microgravity; to determine the effect of microgravity on the actin cytoskeletal organization in gravity-perceiving cells and to study microgravity effects on actin cytoskeleton-related gene expression in plant cells. This investigation allows direct correlation of the results from cytological investigations and gene profiling in order to understand the nature of the actin cytoskeleton in mechanisms of gravity perception. This research focuses on the effects of gravity on basic cellular mechanisms and processes in plants. Improved knowledge of the basic mechanistic processes that will be the focus of this research is vital to develop ways to use plants in extraterrestrial bioregenerative life support systems.

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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 and


  • 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 and Protocols
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 Glutaraldehyde, RNAlater, and Formaldehyde). 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..
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 fluids 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 crew interface.

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Decadal Survey Recommendations

CategoryReference
Plant and Microbial Biology P2
Plant and Microbial Biology P3

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Results/More Information

To investigate how plant development is affected by microgravity, Arabidopsis thaliana seedlings were grown in darkness using the Biological Research in Canisters-Petri Dish Fixation Unit (BRIC-PDFU) hardware flown on the space shuttle. Seedlings from the spaceflight experiment (FL) were then compared to a BRIC-PDFU ground control (GC) and those grown in stand-alone petri dishes (HC). Researchers found that growing plants within the BRIC-PDFU flight hardware led to decreased seed germination and differences in physical and cellular structures in the roots systems of the seedlings. The first major physical difference was observed at the root apex and proximal root where extreme screwing occurred in the flight seedlings compared to slightly skewed roots with the ground controls. Another major difference was the greater amount of adventitious roots (roots formed from the stem or leaves) found on the flight samples. Other effects included the percentage of plants arose from seeds were significantly lower in samples grown in flight hardware (FL, GC) compared to the growing seedlings in just the petri dishes. In addition, the endodermal cells (the deepest cells in the outer layer) were significantly smaller in seedlings grown in the BRIC-PDFU system compared to those in the HC. This change in the shape of endodermal cells indicates alterations in the cell wall and appears to be a true microgravity effect. Since the environmental conditions (e.g., temperature, humidity) were the same for all samples, these results suggest a detrimental effect of growing seedlings in the sealed BRIC-PDFU system. To date, the BRIC-PDFU system has been used in a number of spaceflight experiments in plant biology. However, with these successes, the BRIC-PDFU has some limitations as shown by some results. Thus, in future studies, caution is recommend in the interpretation of results with the BRIC-PDFU system, and the use of an additional ground control to separate space-hardware impacts from true microgravity effects.

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Results Publications

    Johnson CM, Subramanian A, Edelmann RE, Kiss JZ.  Morphometric analyses of petioles of seedlings grown in a spaceflight experiment. Journal of Plant Research. 2015 November; 128(6): 1007-1016. DOI: 10.1007/s10265-015-0749-0.

    Millar KD, Johnson CM, Edelmann RE, Kiss JZ.  An Endogenous Growth Pattern of Roots Is Revealed in Seedlings Grown in Microgravity. Astrobiology. 2011 Oct; 787-797(8): 1-12. DOI: 10.1089/ast.2011.0699. PMID: 21970704.

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Ground Based Results Publications

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ISS Patents

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Related Publications

    Kiss JZ, Brinkmann E, Brillouet C.  Development and Growth of Several Strains of Arabidopsis Seedlings in Microgravity. International Journal of Plant Sciences. 2000; 161: 55-62.

    Kiss JZ, Edelmann RE, Wood PC.  Gravitropism of hypocotyls of wild-type and starch-deficient Arabidopsis seedlings in spaceflight studies. Planta. 1999; 209: 96-103.

    Kiss JZ, Kumar P, Bowman RN, Steele MK, Eodice MT, Correll MJ, Edelmann RE.  Biocompatibility studies in preparation for a spaceflight experiment on plant tropisms (TROPI). Advances in Space Research. 2007; 39(7): 1154-1160. DOI: 10.1016/k.asr.2006.12.017.

    Mullen JL, Correll MJ, Hangarter RP.  Phytochromes A and B mediate red-light-induced positive phototropism in roots. Plant Physiology. 2003; 131: 1411-1417.

    Kiss JZ, Wright JB, Casper T.  Gravitropism in roots of intermediate-starch mutants of Arabidopsis. Plant Physiology. 1996; 97: 237-244.

    Kiss JZ.  Mechanisms of the early phases of plant gravitropism. Critical Reviews in Plant Sciences. 0; 19: 551-573.

    Kiss JZ, Kumar P, Millar KD, Edelmann RE, Correll MJ.  Operations of a spaceflight experiment to investigate plant tropisms. Advances in Space Research. 2009; 44(8): 879-886. DOI: 10.1016/l.asr.2009.06.007.

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Related Websites

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Imagery

image Growth of Arabidopsis seedlings in 60-mm Petri dishes in the dark as in the BRIC-LED system. Seedlings of the Columbia ecotype grown in nutrient agar (A) or on a series of membranes (B). The times indicate days after sowing the seeds in darkness. Image courtesy of J.Z.Kiss.
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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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image Seedlings of Arabidopsis thaliana grown in microgravity in the BRIC system on STS-131. Note the skewing of the root system.
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image NASA Image: JSC2010E057530 - Image of Biological Research In Canisters (BRIC)-16 middeck payload hardware.
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image NASA Image: S135E012237 - Astronaut Rex Walheim works with the Biological Research in Canisters experiment.
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image NASA Image: S135E012252 - Astronaut Rex Walheim poses for a photo with Biological Research in Canisters experiment.
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