Proteomic signature of Arabidopsis cell cultures exposed to magnetically induced hyper- and microgravity environments.

Earth-based microgravity simulation techniques are required due to space research constraints. Using diamagnetic levitation, we exposed Arabidopsis thaliana in vitro callus cultures to environments with different levels of effective gravity and magnetic field strengths (B) simultaneously. The environments included simulated 0 g* at B=10.1 T, an internal 1 g* control (B=16.5 T), and hypergravity (2 g* at B=10.1 T). Furthermore, samples were also exposed to altered gravity environments that were created with mechanical devices, such as the Random Positioning Machine (simulated μg) and the Large Diameter Centrifuge (2 g). We have determined the proteomic signature of cell cultures exposed to these altered-gravity environments by means of the difference gel electrophoresis (DiGE) technique, and we have compared the results with microarray-based transcriptomes from the same samples. The magnetic field itself produced a low number of proteomic alterations, but the combination of gravitational alteration and magnetic field exposure produced synergistic effects on the proteome of plants (the number of significant changes is 3-7 times greater). Tandem mass spectrometry identification of 19 overlapping spots in the different conditions corroborates a major role of abiotic stress and secondary metabolism proteins in the molecular adaptation of plants to unusual environments, including microgravity.

[1]  M. Menges,et al.  Synchronization, transformation, and cryopreservation of suspension-cultured cells. , 2006, Methods in molecular biology.

[2]  K. Guevorkian,et al.  Swimming Paramecium in magnetically simulated enhanced, reduced, and inverted gravity environments , 2006, Proceedings of the National Academy of Sciences.

[3]  H. Engelkamp,et al.  The High Field Magnet Laboratory at Radboud University Nijmegen , 2010 .

[4]  Andrew R Jones,et al.  A DIGE study on the effects of salbutamol on the rat muscle proteome - an exemplar of best practice for data sharing in proteomics , 2011, BMC Research Notes.

[5]  P. Christianen,et al.  The new installation at the Nijmegen High Field Magnet Laboratory , 2004 .

[6]  G. Seidel,et al.  Magnetic levitation-based Martian and Lunar gravity simulator. , 2005, Advances in space research : the official journal of the Committee on Space Research.

[7]  Johannes Madlung,et al.  Changes in the effective gravitational field strength affect the state of phosphorylation of stress-related proteins in callus cultures of Arabidopsis thaliana , 2009 .

[8]  R. Marco,et al.  Microgravity simulation by diamagnetic levitation: effects of a strong gradient magnetic field on the transcriptional profile of Drosophila melanogaster , 2012, BMC Genomics.

[9]  Laurence Eaves,et al.  Effect of magnetically simulated zero-gravity and enhanced gravity on the walk of the common fruitfly† , 2012, Journal of The Royal Society Interface.

[10]  Xiaoping Chen,et al.  Transcriptome profiling of peanut (Arachis hypogaea) gynophores in gravitropic response. , 2013, Functional plant biology : FPB.

[11]  B. Ren,et al.  Magnetic Field Is the Dominant Factor to Induce the Response of Streptomyces avermitilis in Altered Gravity Simulated by Diamagnetic Levitation , 2011, PloS one.

[12]  Shotgun Mass Spectrometry Workflow Combining IEF and LC-MALDI-TOF/TOF , 2010, The protein journal.

[13]  J. Denegre,et al.  Stable magnetic field gradient levitation of Xenopus laevis: toward low-gravity simulation. , 1996, Biophysical journal.

[14]  J. Loon,et al.  The Large Diameter Centrifuge, LDC, for Life and Physical Sciences and Technology , 2008 .

[15]  Wei Sha,et al.  A proteomic approach to analysing responses of Arabidopsis thaliana callus cells to clinostat rotation. , 2006, Journal of experimental botany.

[16]  R. Herranz,et al.  Mechanisms of disruption of meristematic competence by microgravity in Arabidopsis seedlings , 2014, Plant signaling & behavior.

[17]  Julia C. Engelmann,et al.  Transcriptome Analysis in Tardigrade Species Reveals Specific Molecular Pathways for Stress Adaptations , 2012, Bioinformatics and biology insights.

[18]  P. C. Williams,et al.  Magnetic Levitation of MC3T3 Osteoblast Cells as a Ground-Based Simulation of Microgravity , 2009, Microgravity science and technology.

[19]  R. Tournier,et al.  Levitation of organic materials , 1991, Nature.

[20]  Gilbert Gasset,et al.  Spaceflight‐related suboptimal conditions can accentuate the altered gravity response of Drosophila transcriptome , 2010, Molecular ecology.

[21]  Raul Herranz,et al.  Gravitational and magnetic field variations synergize to cause subtle variations in the global transcriptional state of Arabidopsis in vitro callus cultures , 2012, BMC Genomics.

[22]  M. Davey,et al.  Diamagnetic levitation enhances growth of liquid bacterial cultures by increasing oxygen availability , 2010, Journal of The Royal Society Interface.

[23]  Johannes Madlung,et al.  Time-course of changes in amounts of specific proteins upon exposure to hyper-g, 2-D clinorotation, and 3-D random positioning of Arabidopsis cell cultures. , 2007, Journal of experimental botany.

[24]  Yoshio Masuda,et al.  Changes in plant growth processes under microgravity conditions simulated by a three-dimensional clinostat , 1992, The botanical magazine = Shokubutsu-gaku-zasshi.

[25]  Jack J. W. A. van Loon,et al.  Some history and use of the random positioning machine, RPM, in gravity related research , 2007 .

[26]  Eric Beaugnon,et al.  Levitation of water and organic substances in high static magnetic fields , 1991 .

[27]  Michael V Berry,et al.  Of flying frogs and levitrons , 1997 .

[28]  M. Bizzarri,et al.  Gravity sensing by cells: mechanisms and theoretical grounds , 2014, Rendiconti Lincei.

[29]  J. Kuhl,et al.  Differential effects of environment on potato phenylpropanoid and carotenoid expression , 2012, BMC Plant Biology.

[30]  Anne-Marie Faber,et al.  Gel‐based and gel‐free proteomic analysis of Nicotiana tabacum trichomes identifies proteins involved in secondary metabolism and in the (a)biotic stress response , 2011, Proteomics.

[31]  J. Schenck,et al.  Health and Physiological Effects of Human Exposure to Whole‐Body Four‐Tesla Magnetic Fields during MRI , 1992, Annals of the New York Academy of Sciences.

[32]  Jens Hauslage,et al.  Ground-based facilities for simulation of microgravity: organism-specific recommendations for their use, and recommended terminology. , 2013, Astrobiology.

[33]  John Z. Kiss,et al.  Plastid position in Arabidopsis columella cells is similar in microgravity and on a random-positioning machine , 2000, Planta.

[34]  C. Dijkstra,et al.  Bacillus thuringiensis conjugation in simulated microgravity. , 2009, Astrobiology.