Progress 04/16/07 to 05/31/11
OUTPUTS: Outputs. The activities of the past year were focused on the data analyses of the samples returned from the three launches of the TAGES experiments: STS-129, STS-130 STS-131. Dissemination of the information (in addition to publication) includes presentations by both PIs at the Gravitational and Space Biology meeting in 2011 in Santa Claire, CA, at the suborbital meeting in Orlando FL, and at NASA Headquarters. Both PIs are involved with extensive outreach activities in local schools. PARTICIPANTS: Individuals responsible for the Project: PI: Robert J. Ferl and Co-PI: Anna-Lisa Paul Collaborator: Dr. Alain Berinstain at the Canadian Space Agency (PI for the Arthur Clarke Mars Greenhouse project on Devon Island); Dr. Howard Levine at Kennedy Space Center (Science Coordinator for the TAGES experiment at KSC) Training or professional development: Graduate Students during life of the project: John Mayfield, Anne Visscher, Tufan Gokirmak, Joe Collins; undergraduate students: Claire Amalfitano, Lauren Frizzell, Justin Goodwyn. TARGET AUDIENCES: The scientific community and the general community (both adult and student) though the educational outreach activities outlined in a previous section. PROJECT MODIFICATIONS: Not relevant to this project.
PROGRESS: Samples were collected and processed from the experimental subsets launched on STS-129, 130 and 131 in 2009/2010. Two approaches of data collection were used; GFP reporter genes were used to collect gene expression data in real time within GIS (GFP Imaging System) imaging hardware, growth morphology images were also collected, and plants were harvested on orbit to RNAlater for subsequent gene expression profiling with Affymetrix and RNASeq transcriptome analyses. Leaves, hypocotyls and roots were examined independently, revealing that there are a number of organ-specific responses that Arabidopsis employs to respond to the spaceflight environment. Almost a third of the statistically significant, differentially expressed genes encode proteins typically associated with cell wall remodeling, including touch, pathogen response and wounding. Many were dramatically induced (between 5 and 25-fold). In addition, the unique imaging capabilities of the GIS housed in the ABRS unit enabled us to collect morphometric data on the patterns of root growth in the GFP reporter gene plants. These data revealed that there are fundamental differences in root growth and development on orbit, and that two common cultivars of Arabidopsis Wassilewskija (WS) and Columbia (Col-0) are distinct in their responses to spaceflight. This year we also launched a successful parabolic aircraft flight campaign in Houston. The parabolic flight experiment tested new hardware configurations suitable for a transition into the suborbital aircraft platform. IMPACT: The overall goal of our program is to understand fundamental molecular biological responses to extraterrestrial environments. This goal aligns with Fundamental Space Biology goals to understand the impact of spaceflight on biological systems to better enable the exploration imperative. We have used molecular biology and genetically tagged plants as biological monitors of spaceflight and space-related environments with great science return, leading to the development of new biological and hardware tools to study the spaceflight response with even higher fidelity in a wider variety of spaceflight-related environments. Gaining insights into such mechanisms is recognized as fundamental within the decadal study and underpin answers to some of the biggest questions in spaceflight plant biology. Although the analyses are ongoing, our data show that spaceflight induces a touch-wound-remodeling response that is linked to distinct morphometric changes in plant growth and development on orbit. These data provide insights into the fundamental nature of root cell growth patterns, and into the nature of the enhanced susceptibility of plants to pathogens in orbital environments
- Denison F., A-L. Paul, A.K. Zupanska, R.J. Ferl, (2011) 14-3-3 Proteins in Plant Physiology Seminars in Cell and Developmental Biology 22(7): 720-727
- Paul A-L, Zupanska A, Ostrow DT, Zhang Y, Sun Y, Li J-L, Shanker S, Farmerie WG, Amalfitano CE, Ferl RJ. Spaceflight transcriptomes: unique responses to a novel environment (2012) Astrobiology 12(1): 40-56
- Paul A-L and Ferl RJ (2011) Using green fluorescent protein (GFP) reporter genes in RNAlater fixed tissue Gravitational and Space Biology 25(1): 40-43
- Paul A-L, Manak MS, Mayfield JD, Reyes MF, Gurley WB, and Ferl RJ. (2011) Parabolic flight induces changes in gene expression patterns in Arabidopsis thaliana. Astrobiology 2011 Oct;11(8):743-58
- Ferl RJ, Zupanska A, Spinale A, Reed D, Manning-Roach S, Guerra G, Cox D, Paul A-L (2011) Performance of KSC Fixation Tubes with RNALater for orbital experiments: a case study in ISS operations for molecular biology. Advances in Space Research 48: 199-206
Progress 10/01/09 to 09/30/10
OUTPUTS: Activities. This academic year, the Payload Verification Test (PVT) was completed, along with several additional tests and experiments in preparation for the launch of the first component of the TAGES experiment (Run 1) on November 16th. The Imaging Hardware continued to undergo significant improvements right up to launch. In addition, insights gained during flight operations contributed to further improvements to the hardware and procedures. Most of the year's effort was focused on the execution and analyses of experiments leading up to the 11-16-2009 launch of the first leg of the TAGES flight experiment. However, the full report of Flight operations and experimental results will be presented in the final report. Events Both PIs presented talks at the Gravitational and Space Biology meeting in Washington DC, 11-2009. Dissemination- both PIs are involved with extensive outreach activities. This year the PIs were asked to help prepare and article describing their space biology research for Explore magazine, and Paul helped to create a series of science videos for lesson plans related to the concept of researching and growing plants in spaceflight and novel planetary environments. These are posted on the UF Genetics web site and on You-Tube. Paul also was invited to present at the career fair for the national organization of Minorities in Agriculture, Natural Resources and Related Sciences (MANRRS) and to the Institute for Learning in Retirement. PARTICIPANTS: Individuals responsible for the Project: PI: Robert J. Ferl and Co-PI: Anna-Lisa Paul Collaborator: Dr. Alain Berinstain at the Canadian Space Agency (PI for the Arthur Clarke Mars Greenhouse project on Devon Island); Dr. Howard Levine at Kennedy Space Center (Science Coordinator for the TAGES experiment at KSC) Training or professional development: Graduate Students: Anne Visscher, Tufan Gokirmak, Joe Collins; undergraduate student Claire Amalfitano and Lauren Frizzell. TARGET AUDIENCES: The scientific community and the general community (both adult and student) though the educational outreach activities outlined in a previous section. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
PROGRESS: This academic year, the Payload Verification Test (PVT) was completed, along with several additional tests and experiments in preparation for the launch of the the TAGES experiment in November, 2009, February 2010 and April 2010. The Imaging Hardware continued to undergo significant improvements right up to launch. In addition, insights gained during flight operations contributed to further improvements to the hardware and procedures. Most of the year's effort was focused on the execution and analyses of experiments leading up to the 11-16-2009 launch of the first leg of the TAGES flight experiment. Additional to the PVT, several experiments were conducted to prepare for flight operations, including exhaustive testing of the TAGES Green Fluorescent Protein Imaging system (GIS), a series of vibration tests to mimic launch environments and a 30 day functional test to conduct a mock flight scenario. There were three separate launches of the TAGES experiment in November, February and April. An extraordinary amount of time and resources went into preparation of the biology and hardware to support these flights. All were successfully launched and returned. The data analyses were begun in May and are still underway. IMPACT: The data from the Flight experiments have demonstrated that the spaceflight hardware developed for TAGES worked well and will have wide application in the spaceflight community. The gene expression data under analysis suggest that the spaceflight environment can be assayed both through telemetric collections of data with fluorescent reporter gene technology as well as traditional biochemical / molecular genetic techniques that require sample return. Data from both approaches indicate that the spaceflight causes changes in the gene expression patterns of plants exposed top this novel environment.
- Bamsey M, Berinstain A, Graham T, Neron P, S. B, Giroux R, Ferl RJ, Paul A-L, Dixon MA (2009) Developing strategies for automated remote plant production systems: Environmental control and monitoring of the Arthur Clarke Mars Greenhouse in the Canadian High Arctic. Advances in Space Research 44: 1367-1381.
- Visscher AM, Paul AL, Kirst M, Alling AK, Silverstone S, Nechitailo G, Nelson M, Dempster WF, Van Thillo M, Allen JP, Ferl RJ (2009) Effects of a spaceflight environment on heritable changes in wheat gene expression. Astrobiology 9: 359-367.
- Ferl RJ, Paul AL (2010) Lunar Plant Biology- A Review the Apollo Era. Astrobiology 10:261-274
- Visscher AM, Paul AL, Kirst M, Guy C, Schuerger A, Ferl RJ (2010) Growth performance and root transcriptome remodeling of Arabidopsis in response to Mars-like levels of magnesium sulfate. PLoS1 5:e12348
- Gokirmak T, Paul AL, Ferl RJ (2010) Plant phosphopeptide-binding proteins as signaling mediators. Curr Opin Plant Biol 13: 527-532
Progress 10/01/07 to 09/30/08
OUTPUTS: ACTIVITIES in the first year of the grant period (10/1/2007 - 9/30/2008)include three significant off-site experiments. November 2007: the Science Verification Test (SVT) for the flight experiment was conducted at Kennedy Space Center. This test evaluates the technical readiness of the biological systems, experimental approaches and hardware interface designed for the eventual deployment of the Flight experiment. May 2008: a parabolic flight experiment at Johnson Space Center in Houston to evaluate the biological effects of an analog spaceflight environment on Arabidopsis plants. The focus of this experiment was to evaluate genome-wide changes in gene expression patterns and gather tissue-specific data on processes associated with the transduction of auxin mediated signaling. July 2008: experiments in a planetary analog site (Haughton Crater in the Canadian high arctic) to evaluate the deployment of remote imaging hardware in the autonomous Arthur Clarke Mars Greenhouse (ACMG) managed by the Canadian Space Agency, and also to collect and evaluate local impact crater materials (breccias) to use as analogs for in situ utilization of local resources on planetary expeditions to the moon or mars. During the course of these experiments, as well as the subsequent analyses of materials and process in our home laboratory, we have involved graduate and undergraduate students, as well as incorporated materials and results into the Space Biology class (HOS4905 / PCB6937) that Ferl and Paul are teaching in the Fall Semester. EVENTS include having presentations at the 37th COSPAR Scientific Assembly in early July and the NASA Lunar Science Institute (NLSI) conference in late July. DISSEMINATION - both PIs are involved with extensive outreach activities. This year the PIs were asked to help prepare and article describing their space biology research for Explore magazine, and Paul helped to create a series of science videos for lesson plans related to the concept of researching and growing plants in spaceflight and novel planetary environments. These are posted on the UF Genetics web site and on You-Tube. Paul also was invited to present to the Florida Association of Agricultural Educators (FAAE) Tech Update. Her participation included a presentation (Growing plants off-planet - What do we need to know before we go), the live demonstration of GFP biosensor plants with GFP fluorescent imaging hardware, and the development of education kits to grow arabidopsis plants in the classroom. PARTICIPANTS: PI: Robert J. Ferl Co-PI: Anna-Lisa Paul. Graduate Students: Anne Visscher and Tufan Gokirmak. Collaborator at the Canadian Space Agency: Alain Berinstain (PI for the Arthur Clarke Mars Greenhouse project on Devon Island). TARGET AUDIENCES: The scientific community and the general community (both adult and student) though the educational outreach activities outlined in a previous section. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
PROGRESS: The second generation of TAGES GFP imager (TIS-II) was tested for remote deployment to mimic aspects of the collection of telemetric data from another planetary surface. We deployed the GFP imaging system within the Arthur Clarke Mars Greenhouse (ACMG) an autonomously operated greenhouse located within the Haughton Mars Project in the on Devon Island in the Canadian high arctic. Results both demonstrate the applicability of the fundamental GFP biosensor technology and highlight the difficulties in collecting and managing telemetric data from challenging deployment environments. Parabolic flights were conducted on the NASA C-9 aircraft to explore aspects of signal transduction in arabidopsis. Microarray data from a previous KC-135 experiment indicated that several auxin-associated genes are affected by the parabolic flight environment. Two were chosen to test the efficacy of biosensors to report tissue-specific patterns of gene expression of early auxin signaling. Pin4 encodes an auxin efflux mediator and Arf7 encodes a transcription factor that binds to the Auxin Responsive element. Plants were harvested at three time points during the C-9 flight: before flight as Ground Control at parabola 20 and parabola 40. The experiment was conducted twice. The histochemical staining patterns of the Pin4 and Arf7 GUS reporters indicates that both of these auxin signaling genes are differentially-regulated in a tissue-specific manner in response to parabolic flight environments. IMPACT: The data from the experiment and spaceflight hardware development have resulted in a design that should successfully grow plants on the space station and analyze gene expression of plants telemetrically. The analog environment experiments corroborate that the Imaging Hardware under development for Flight application can also be adapted for remote sensing applications in platforms such as planetary landers and suit-case deployment by astronauts returning to the moon. The C-9 experiments demonstrate that applicability of parabolic flight as an analog test environment for spaceflight applications and provide insight into the signal transduction associated with the role of auxin in early gravity perception. These accomplishments set the stage for experiments that explore the limits of agricultural biology within the solar system and extend knowledge that might allow the production of food and air/water recycling in extraterrestrial habitats.
- Paul, A-L., Bamsey, M., Berinstain, A., Braham. S., Neron, P., Murdoch T., Graham, T. and R. J. Ferl (2008). Deployment of a Prototype Plant GFP Imager at the Arthur Clarke Mars Greenhouse of the Haughton Mars Project. Sensors, 8: 2762-2773.