Microgravity Induces Changes in Microsome-Associated Proteins of Arabidopsis Seedlings Grown on Board the International Space Station

The “GENARA A” experiment was designed to monitor global changes in the proteome of membranes of Arabidopsis thaliana seedlings subjected to microgravity on board the International Space Station (ISS). For this purpose, 12-day-old seedlings were grown either in space, in the European Modular Cultivation System (EMCS) under microgravity or on a 1 g centrifuge, or on the ground. Proteins associated to membranes were selectively extracted from microsomes and identified and quantified through LC-MS-MS using a label-free method. Among the 1484 proteins identified and quantified in the 3 conditions mentioned above, 80 membrane-associated proteins were significantly more abundant in seedlings grown under microgravity in space than under 1 g (space and ground) and 69 were less abundant. Clustering of these proteins according to their predicted function indicates that proteins associated to auxin metabolism and trafficking were depleted in the microsomal fraction in µg space conditions, whereas proteins associated to stress responses, defence and metabolism were more abundant in µg than in 1 g indicating that microgravity is perceived by plants as a stressful environment. These results clearly indicate that a global membrane proteomics approach gives a snapshot of the cell status and its signaling activity in response to microgravity and highlight the major processes affected.

[1]  Mari L. Salmi,et al.  Changes in gravity rapidly alter the magnitude and direction of a cellular calcium current , 2011, Planta.

[2]  O. Monje,et al.  Microgravity effects on leaf morphology, cell structure, carbon metabolism and mRNA expression of dwarf wheat , 2006, Planta.

[3]  Influence of long term exposure to space flight on tomato seeds , 2005 .

[4]  B. Gunning,et al.  Structure of cortical microtubule arrays in plant cells , 1978, The Journal of cell biology.

[5]  T. Ziv,et al.  Proteomics of the response of Arabidopsis thaliana to infection with Alternaria brassicicola. , 2010, Journal of proteomics.

[6]  Zhaoyuan Fang,et al.  Phosphate signaling in Arabidopsis and Oryza sativa , 2009 .

[7]  J. Adamec,et al.  ABCB19/PGP19 stabilises PIN1 in membrane microdomains in Arabidopsis. , 2009, The Plant journal : for cell and molecular biology.

[8]  Yukiko Nakamura,et al.  Cell wall changes involved in the automorphic curvature of rice coleoptiles under microgravity conditions in space , 2004, Journal of Plant Research.

[9]  P. Naur,et al.  CYP83B1 Is the Oxime-metabolizing Enzyme in the Glucosinolate Pathway in Arabidopsis * , 2001, The Journal of Biological Chemistry.

[10]  Daowen Wang,et al.  The Arabidopsis Purple Acid Phosphatase AtPAP10 Is Predominantly Associated with the Root Surface and Plays an Important Role in Plant Tolerance to Phosphate Limitation1[W][OA] , 2011, Plant Physiology.

[11]  Robert J Ferl,et al.  Fundamental plant biology enabled by the space shuttle. , 2013, American journal of botany.

[12]  P. Masson Root gravitropism , 1995, BioEssays : news and reviews in molecular, cellular and developmental biology.

[13]  P. Benfey,et al.  Genetic evidence that the endodermis is essential for shoot gravitropism in Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.

[14]  S. Bak,et al.  The involvement of two p450 enzymes, CYP83B1 and CYP83A1, in auxin homeostasis and glucosinolate biosynthesis. , 2001, Plant physiology.

[15]  P. Masson,et al.  Multiple roles for membrane-associated protein trafficking and signaling in gravitropism , 2012, Front. Plant Sci..

[16]  T. Hoson,et al.  Growth and cell wall changes in rice roots during spaceflight , 2003, Plant and Soil.

[17]  D. Blaudez,et al.  Structure and Expression Profile of the Phosphate Pht1 Transporter Gene Family in Mycorrhizal Populus trichocarpa1[W] , 2011, Plant Physiology.

[18]  A. D. Krikorian,et al.  Growth and Photosynthetic Responses of Wheat Plants Grown in Space , 1996, Plant physiology.

[19]  R. Wheeler,et al.  Gravitropism in higher plant shoots. IV. Further studies on participation of ethylene. , 1986, Plant physiology.

[20]  G. Perbal,et al.  A polarized cell: the root statocyte. , 2003, Physiologia plantarum.

[21]  B S Spooner,et al.  Impact of altered gravity on aspects of cell biology. , 1994, International review of cytology.

[22]  W. Lukowitz,et al.  The gravitropism defective 2 Mutants of Arabidopsis Are Deficient in a Protein Implicated in Endocytosis in Caenorhabditis elegans1[w] , 2004, Plant Physiology.

[23]  M. Hamberg,et al.  Oxylipins Produced by the 9-Lipoxygenase Pathway in Arabidopsis Regulate Lateral Root Development and Defense Responses through a Specific Signaling Cascade[W] , 2007, The Plant Cell Online.

[24]  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.

[25]  Jiří Friml,et al.  Intracellular trafficking and proteolysis of the Arabidopsis auxin-efflux facilitator PIN2 are involved in root gravitropism , 2006, Nature Cell Biology.

[26]  Daowen Wang,et al.  Biochemical and molecular characterization of AtPAP12 and AtPAP26: the predominant purple acid phosphatase isozymes secreted by phosphate-starved Arabidopsis thaliana. , 2010, Plant, cell & environment.

[27]  T. Hoson,et al.  New aspects of gravity responses in plant cells. , 2003, International review of cytology.

[28]  F. Salisbury,et al.  Gravitropism in higher plant shoots. V. Changing sensitivity to auxin. , 1988, Plant physiology.

[29]  D. Weijers,et al.  The PINOID protein kinase regulates organ development in Arabidopsis by enhancing polar auxin transport. , 2001, Development.

[30]  M. Morita,et al.  A SNARE complex containing SGR3/AtVAM3 and ZIG/VTI11 in gravity-sensing cells is important for Arabidopsis shoot gravitropism , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[31]  K. Palme,et al.  Jasmonate modulates endocytosis and plasma membrane accumulation of the Arabidopsis PIN2 protein. , 2011, The New phytologist.

[32]  J. Dinneny,et al.  Tools for high-spatial and temporal-resolution analysis of environmental responses in plants , 2010, Biotechnology Letters.

[33]  Odile Burlet-Schiltz,et al.  Mascot File Parsing and Quantification (MFPaQ), a New Software to Parse, Validate, and Quantify Proteomics Data Generated by ICAT and SILAC Mass Spectrometric Analyses , 2007, Molecular & Cellular Proteomics.

[34]  J E Mullet,et al.  A chloroplast lipoxygenase is required for wound-induced jasmonic acid accumulation in Arabidopsis. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[35]  J. Aarrouf,et al.  Changes in hormonal balance and meristematic activity in primary root tips on the slowly rotating clinostat and their effect on the development of the rapeseed root system. , 1999, Physiologia plantarum.

[36]  Ziding Zhang,et al.  MDP25, A Novel Calcium Regulatory Protein, Mediates Hypocotyl Cell Elongation by Destabilizing Cortical Microtubules in Arabidopsis[C][W][OA] , 2011, Plant Cell.

[37]  E. Blancaflor,et al.  Regulation of plant gravity sensing and signaling by the actin cytoskeleton. , 2013, American journal of botany.

[38]  Klaus Palme,et al.  A PINOID-Dependent Binary Switch in Apical-Basal PIN Polar Targeting Directs Auxin Efflux , 2004, Science.

[39]  K. Hirschi,et al.  Vacuolar CAX1 and CAX3 Influence Auxin Transport in Guard Cells via Regulation of Apoplastic pH1[W][OA] , 2012, Plant Physiology.

[40]  H. Fukaki,et al.  SGR1, SGR2, and SGR3: Novel Genetic Loci Involved in Shoot Gravitropism in Arabidopsis thaliana , 1996, Plant physiology.

[41]  B. Pickard,et al.  Gravitropism in a starchless mutant of Arabidopsis: implications for the starch-statolith theory of gravity sensing. , 1989, Planta.

[42]  B. Usadel,et al.  The Interconversion of UDP-Arabinopyranose and UDP-Arabinofuranose Is Indispensable for Plant Development in Arabidopsis[C][W][OA] , 2011, Plant Cell.

[43]  F. Ausubel,et al.  An Arabidopsis thaliana Lipoxygenase Gene Can Be Induced by Pathogens, Abscisic Acid, and Methyl Jasmonate , 1993, Plant physiology.

[44]  C. Wasternack,et al.  Jasmonate biosynthesis and the allene oxide cyclase family of Arabidopsis thaliana , 2003, Plant Molecular Biology.

[45]  Eugénie Carnero-Diaz,et al.  The dose-response curve of the gravitropic reaction: a re-analysis. , 2002, Physiologia plantarum.

[46]  G. Perbal,et al.  Perception of gravity in the lentil root , 2004, Naturwissenschaften.

[47]  F. Ausubel,et al.  The Apoplastic Oxidative Burst Peroxidase in Arabidopsis Is a Major Component of Pattern-Triggered Immunity[W][OA] , 2012, Plant Cell.

[48]  E. Bayer,et al.  Membrane rafts in plant cells. , 2010, Trends in plant science.

[49]  Richard E. Edelmann,et al.  Transcriptome analyses of Arabidopsis thaliana seedlings grown in space: implications for gravity-responsive genes , 2013, Planta.

[50]  K. Ljung,et al.  The SUR2 gene of Arabidopsis thaliana encodes the cytochrome P450 CYP83B1, a modulator of auxin homeostasis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[51]  Erich Kombrink,et al.  A New Type of Peroxisomal Acyl-Coenzyme A Synthetase from Arabidopsis thaliana Has the Catalytic Capacity to Activate Biosynthetic Precursors of Jasmonic Acid* , 2005, Journal of Biological Chemistry.

[52]  M. Maeshima,et al.  A hydrophilic cation‐binding protein of Arabidopsis thaliana, AtPCaP1, is localized to plasma membrane via N‐myristoylation and interacts with calmodulin and the phosphatidylinositol phosphates PtdIns(3,4,5)P3 and PtdIns(3,5)P2 , 2008, The FEBS journal.

[53]  T. Hoson,et al.  Cell wall-bound peroxidase activity and lignin formation in azuki bean epicotyls grown under hypergravity conditions. , 2009, Journal of plant physiology.

[54]  P. Masson,et al.  Adenosine Kinase Modulates Root Gravitropism and Cap Morphogenesis in Arabidopsis1[W][OA] , 2006, Plant Physiology.

[55]  B. Bartel,et al.  Redundancy as a way of life - IAA metabolism. , 1999, Current opinion in plant biology.

[56]  T. McNellis,et al.  A Humidity-Sensitive Arabidopsis Copine Mutant Exhibits Precocious Cell Death and Increased Disease Resistance , 2001, The Plant Cell Online.

[57]  R. Yamamoto,et al.  Effect of simulated microgravity on auxin polar transport in inflorescence axis of Arabidopsis thaliana. , 1995, Uchu Seibutsu Kagaku.

[58]  R. Offringa,et al.  PINOID-Mediated Signaling Involves Calcium-Binding Proteins , 2003, Plant Physiology.

[59]  Ben Scheres,et al.  Polar PIN Localization Directs Auxin Flow in Plants , 2006, Science.

[60]  K. Fukui,et al.  Growth and Development, and Auxin Polar Transport in Higher Plants under Microgravity Conditions in Space: BRIC-AUX on STS-95 Space Experiment , 1999, Journal of Plant Research.

[61]  R. Herranz,et al.  Exploration of plant growth and development using the European Modular Cultivation System facility on the International Space Station. , 2014, Plant biology.

[62]  Dominique Driss-Ecole,et al.  Mechanotransduction in gravisensing cells. , 2003, Trends in plant science.

[63]  Zachary B. Abrams,et al.  A proteomics approach identifies novel proteins involved in gravitropic signal transduction. , 2013, American journal of botany.

[64]  K. Hasenstein Gravisensing in plants and fungi. , 1999, Advances in Space Research.

[65]  T. Hoson,et al.  Changes in levels of cell wall constituents in wheat seedlings grown under continuous hypergravity conditions , 2005 .

[66]  D. Galbraith,et al.  CYP83B1, a Cytochrome P450 at the Metabolic Branch Point in Auxin and Indole Glucosinolate Biosynthesis in Arabidopsis , 2001, Plant Cell.

[67]  R. Offringa,et al.  Plant evolution: AGC kinases tell the auxin tale. , 2007, Trends in plant science.

[68]  H. Fukaki,et al.  Mutations in the SGR4, SGR5 and SGR6 loci of Arabidopsis thaliana alter the shoot gravitropism. , 1997, Plant & cell physiology.

[69]  P. Staswick The Tryptophan Conjugates of Jasmonic and Indole-3-Acetic Acids Are Endogenous Auxin Inhibitors1[W][OA] , 2009, Plant Physiology.

[70]  B. Jeune,et al.  Lentil root statoliths reach a stable state in microgravity , 2000, Planta.

[71]  John Z. Kiss,et al.  Amyloplasts are necessary for full gravitropic sensitivity in roots of Arabidopsis thaliana , 2004, Planta.

[72]  Robert J Ferl,et al.  Spaceflight transcriptomes: unique responses to a novel environment. , 2012, Astrobiology.

[73]  P. Masson,et al.  Gravitropism in higher plants. , 1999, Plant physiology.

[74]  R. Hampp,et al.  Hyper-gravity effects on the Arabidopsis transcriptome. , 2003, Physiologia plantarum.

[75]  P. Dayanandan Gravitational biology and space life sciences: Current status and implications for the Indian space programme , 2011, Journal of Biosciences.

[76]  H. Xue,et al.  Arabidopsis phosphatidylinositol monophosphate 5-kinase 2 is involved in root gravitropism through regulation of polar auxin transport by affecting the cycling of PIN proteins , 2011, Cell Research.

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

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

[79]  G. Fink,et al.  The BON/CPN gene family represses cell death and promotes cell growth in Arabidopsis. , 2006, The Plant journal : for cell and molecular biology.

[80]  K. Okada,et al.  phot1 and phot2 mediate blue light-induced transient increases in cytosolic Ca2+ differently in Arabidopsis leaves , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[81]  J. Leplé,et al.  Proteome analysis of apical and basal regions of poplar stems under gravitropic stimulation. , 2009, Physiologia plantarum.

[82]  D. Driss-Ecole,et al.  [Microgravity and root gravitropism]. , 1993, Acta botanica Gallica : bulletin de la Societe botanique de France.