The Reductase Activity of the Arabidopsis Caleosin RESPONSIVE TO DESSICATION20 Mediates Gibberellin-Dependent Flowering Time, Abscisic Acid Sensitivity, and Tolerance to Oxidative Stress1[W]

A peroxygenase controls floral transition and tolerance to oxidative stress. Contrasting with the wealth of information available on the multiple roles of jasmonates in plant development and defense, knowledge about the functions and the biosynthesis of hydroxylated oxylipins remains scarce. By expressing the caleosin RESPONSIVE TO DESSICATION20 (RD20) in Saccharomyces cerevisiae, we show that the recombinant protein possesses an unusual peroxygenase activity with restricted specificity toward hydroperoxides of unsaturated fatty acid. Accordingly, Arabidopsis (Arabidopsis thaliana) plants overexpressing RD20 accumulate the product 13-hydroxy-9,11,15-octadecatrienoic acid, a linolenate-derived hydroxide. These plants exhibit elevated levels of reactive oxygen species (ROS) associated with early gibberellin-dependent flowering and abscisic acid hypersensitivity at seed germination. These phenotypes are dependent on the presence of active RD20, since they are abolished in the rd20 null mutant and in lines overexpressing RD20, in which peroxygenase was inactivated by a point mutation of a catalytic histidine residue. RD20 also confers tolerance against stress induced by Paraquat, Rose Bengal, heavy metal, and the synthetic auxins 1-naphthaleneacetic acid and 2,4-dichlorophenoxyacetic acid. Under oxidative stress, 13-hydroxy-9,11,15-octadecatrienoic acid still accumulates in RD20-overexpressing lines, but this lipid oxidation is associated with reduced ROS levels, minor cell death, and delayed floral transition. A model is discussed where the interplay between fatty acid hydroxides generated by RD20 and ROS is counteracted by ethylene during development in unstressed environments.

[1]  Kazuki Saito,et al.  Leaf Oil Body Functions as a Subcellular Factory for the Production of a Phytoalexin in Arabidopsis1[W] , 2013, Plant Physiology.

[2]  W. Kuo,et al.  The Arabidopsis ETHYLENE RESPONSE FACTOR1 Regulates Abiotic Stress-Responsive Gene Expression by Binding to Different cis-Acting Elements in Response to Different Stress Signals1[W][OA] , 2013, Plant Physiology.

[3]  I. Baldwin,et al.  High levels of jasmonic acid antagonize the biosynthesis of gibberellins and inhibit the growth of Nicotiana attenuata stems. , 2013, The Plant journal : for cell and molecular biology.

[4]  I. Baldwin,et al.  Nicotiana attenuata α-DIOXYGENASE1 through its production of 2-hydroxylinolenic acid is required for intact plant defense expression against attack from Manduca sexta larvae. , 2012, The New phytologist.

[5]  M. Fauconnier,et al.  A non‐canonical caleosin from Arabidopsis efficiently epoxidizes physiological unsaturated fatty acids with complete stereoselectivity , 2012, The FEBS journal.

[6]  Kevin W Eliceiri,et al.  NIH Image to ImageJ: 25 years of image analysis , 2012, Nature Methods.

[7]  X. Deng,et al.  Plant hormone jasmonate prioritizes defense over growth by interfering with gibberellin signaling cascade , 2012, Proceedings of the National Academy of Sciences.

[8]  L. Soubigou-Taconnat,et al.  Carotenoid oxidation products are stress signals that mediate gene responses to singlet oxygen in plants , 2012, Proceedings of the National Academy of Sciences.

[9]  M. Hamberg,et al.  Antagonistic role of 9-lipoxygenase-derived oxylipins and ethylene in the control of oxidative stress, lipid peroxidation and plant defence. , 2011, The Plant journal : for cell and molecular biology.

[10]  H. Khalil,et al.  Heterotrimeric Gα subunit from wheat (Triticum aestivum), GA3, interacts with the calcium-binding protein, Clo3, and the phosphoinositide-specific phospholipase C, PI-PLC1 , 2011, Plant Molecular Biology.

[11]  J. Jeung,et al.  A stress-responsive caleosin-like protein, AtCLO4, acts as a negative regulator of ABA responses in Arabidopsis. , 2011, Plant and Cell Physiology.

[12]  Alain Vavasseur,et al.  RD20, a stress-inducible caleosin, participates in stomatal control, transpiration and drought tolerance in Arabidopsis thaliana. , 2010, Plant & cell physiology.

[13]  J. Tzen,et al.  Caleosin serves as the major structural protein as efficient as oleosin on the surface of seed oil bodies , 2010, Plant signaling & behavior.

[14]  B. Hwang,et al.  The Pepper 9-Lipoxygenase Gene CaLOX1 Functions in Defense and Cell Death Responses to Microbial Pathogens1[C][W][OA] , 2009, Plant Physiology.

[15]  D. Murphy,et al.  Roles of a membrane-bound caleosin and putative peroxygenase in biotic and abiotic stress responses in Arabidopsis. , 2009, Plant physiology and biochemistry : PPB.

[16]  I. Feussner,et al.  Oxylipins: structurally diverse metabolites from fatty acid oxidation. , 2009, Plant physiology and biochemistry : PPB.

[17]  D. T. Britto,et al.  K+ transport in plants: physiology and molecular biology. , 2009, Journal of plant physiology.

[18]  C. le Bon,et al.  Caleosin of Arabidopsis thaliana : Effect of Calcium on Functional and Structural Properties. , 2008, Journal of agricultural and food chemistry.

[19]  N. Provart,et al.  An extensive (co-)expression analysis tool for the cytochrome P450 superfamily in Arabidopsis thaliana , 2008, BMC Plant Biology.

[20]  F. Zhou,et al.  The developmental transition to flowering in Arabidopsis is associated with an increase in leaf chloroplastic lipoxygenase activity. , 2008, Plant science : an international journal of experimental plant biology.

[21]  S. Mongrand,et al.  Genome-Wide Annotation of Remorins, a Plant-Specific Protein Family: Evolutionary and Functional Perspectives1[W] , 2007, Plant Physiology.

[22]  D. Van Der Straeten,et al.  The plant stress hormone ethylene controls floral transition via DELLA-dependent regulation of floral meristem-identity genes , 2007, Proceedings of the National Academy of Sciences.

[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]  J. Garin,et al.  Plant Seed Peroxygenase Is an Original Heme-oxygenase with an EF-hand Calcium Binding Motif* , 2006, Journal of Biological Chemistry.

[25]  F. Van Breusegem,et al.  Reactive oxygen species as signals that modulate plant stress responses and programmed cell death , 2006, BioEssays : news and reviews in molecular, cellular and developmental biology.

[26]  M. Crespi,et al.  Cross-talk between ethylene and drought signalling pathways is mediated by the sunflower Hahb-4 transcription factor. , 2006, The Plant journal : for cell and molecular biology.

[27]  J. Browse,et al.  A role for caleosin in degradation of oil-body storage lipid during seed germination. , 2006, The Plant journal : for cell and molecular biology.

[28]  Patrick Achard,et al.  Integration of Plant Responses to Environmentally Activated Phytohormonal Signals , 2006, Science.

[29]  Gerhard Leubner-Metzger,et al.  Plant hormone interactions during seed dormancy release and germination , 2005, Seed Science Research.

[30]  M. Hamberg,et al.  Evaluation of the Antimicrobial Activities of Plant Oxylipins Supports Their Involvement in Defense against Pathogens1[W] , 2005, Plant Physiology.

[31]  D. Heiman,et al.  Salicylate activity. 1. Protection of plants from paraquat injury. , 2005, Journal of agricultural and food chemistry.

[32]  D. Inzé,et al.  Fatty Acid Hydroperoxides and H2O2 in the Execution of Hypersensitive Cell Death in Tobacco Leaves1[w] , 2005, Plant Physiology.

[33]  Alan M. Jones,et al.  Different Signaling and Cell Death Roles of Heterotrimeric G Protein α and β Subunits in the Arabidopsis Oxidative Stress Response to Ozonew⃞ , 2005, The Plant Cell Online.

[34]  T. Aung,et al.  Stress-responsive α-dioxygenase expression in tomato roots , 2005 .

[35]  Thomas Girke,et al.  The Vegetative Vacuole Proteome of Arabidopsis thaliana Reveals Predicted and Unexpected Proteinsw⃞ , 2004, The Plant Cell Online.

[36]  T. Douki,et al.  The upstream oxylipin profile of Arabidopsis thaliana: a tool to scan for oxidative stresses. , 2004, The Plant journal : for cell and molecular biology.

[37]  Kazuo Shinozaki,et al.  A dehydration-induced NAC protein, RD26, is involved in a novel ABA-dependent stress-signaling pathway. , 2004, The Plant journal : for cell and molecular biology.

[38]  M. Hamberg,et al.  Activation of the fatty acid alpha-dioxygenase pathway during bacterial infection of tobacco leaves. Formation of oxylipins protecting against cell death. , 2003, The Journal of biological chemistry.

[39]  Ivo Feussner,et al.  The lipoxygenase pathway. , 2003, Annual review of plant biology.

[40]  Cornelia Göbel,et al.  Rapid Induction of Distinct Stress Responses after the Release of Singlet Oxygen in Arabidopsis Online version contains Web-only data. Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.014662. , 2003, The Plant Cell Online.

[41]  M. Hamberg,et al.  Oxylipin profiling in pathogen-infected potato leaves. , 2002, Biochimica et biophysica acta.

[42]  K. Patel,et al.  The Brassica napus calcium-binding protein, caleosin, has distinct endoplasmic reticulum- and lipid body-associated isoforms , 2001 .

[43]  H. Nielsen,et al.  Caleosins: Ca2+-binding proteins associated with lipid bodies , 2000, Plant Molecular Biology.

[44]  Brown,et al.  The developmental transition to flowering represses ascorbate peroxidase activity and induces enzymatic lipid peroxidation in leaf tissue in Arabidopsis thaliana. , 2000, Plant science : an international journal of experimental plant biology.

[45]  K. Yamaguchi-Shinozaki,et al.  An Arabidopsis gene encoding a Ca2+-binding protein is induced by abscisic acid during dehydration. , 2000, Plant & cell physiology.

[46]  J. Giraudat,et al.  Interactions between Abscisic Acid and Ethylene Signaling Cascades , 2000, Plant Cell.

[47]  P. McCourt,et al.  Regulation of Abscisic Acid Signaling by the Ethylene Response Pathway in Arabidopsis , 2000, Plant Cell.

[48]  M. Koornneef,et al.  Gibberellin requirement for Arabidopsis seed germination is determined both by testa characteristics and embryonic abscisic acid. , 2000, Plant physiology.

[49]  J. Blein,et al.  Involvement of Lipoxygenase-dependent Production of Fatty Acid Hydroperoxides in the Development of the Hypersensitive Cell Death induced by Cryptogein on Tobacco Leaves* , 1999, The Journal of Biological Chemistry.

[50]  M. Hamberg,et al.  alpha-oxidation of fatty acids in higher plants. Identification of a pathogen-inducible oxygenase (piox) as an alpha-dioxygenase and biosynthesis of 2-hydroperoxylinolenic acid. , 1999, The Journal of biological chemistry.

[51]  J. Ecker,et al.  EIN2, a bifunctional transducer of ethylene and stress responses in Arabidopsis. , 1999, Science.

[52]  T. Thomas,et al.  ATS1 and ATS3: two novel embryo-specific genes in Arabidopsis thaliana , 1999, Plant Molecular Biology.

[53]  S. Clough,et al.  Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.

[54]  E. Blée,et al.  Phytooxylipins and plant defense reactions. , 1998, Progress in lipid research.

[55]  David B. Collinge,et al.  Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley—powdery mildew interaction , 1997 .

[56]  K. Shinozaki,et al.  Correlation between the induction of a gene for delta 1-pyrroline-5-carboxylate synthetase and the accumulation of proline in Arabidopsis thaliana under osmotic stress. , 1995, The Plant journal : for cell and molecular biology.

[57]  M. Thomashow,et al.  The 5′-region of Arabidopsis thaliana cor15a has cis-acting elements that confer cold-, drought- and ABA-regulated gene expression , 1994, Plant Molecular Biology.

[58]  L. Marnett,et al.  Mechanism of reaction of fatty acid hydroperoxides with soybean peroxygenase. , 1993, The Journal of biological chemistry.

[59]  R. N. Wilson,et al.  Gibberellin Is Required for Flowering in Arabidopsis thaliana under Short Days. , 1992, Plant physiology.

[60]  K. Shinozaki,et al.  Molecular Cloning and Characterization of 9 cDNAs for Genes That Are Responsive to Desiccation in Arabidopsis thaliana: SequenceAnalysis of One cDNA Clone That Encodes a Putative Transmembrane Channel Protein , 1992 .

[61]  M. Hamberg.  Regio- and stereochemical analysis of trihydroxyoctadecenoic acids derived from linoleic acid 9- and 13-hydroperoxides , 1991, Lipids.

[62]  M. Hamberg,et al.  Hydroperoxide-dependent epoxidation of unsaturated fatty acids in the broad bean (Vicia faba L.). , 1990, Archives of biochemistry and biophysics.

[63]  F. Schuber,et al.  Efficient epoxidation of unsaturated fatty acids by a hydroperoxide-dependent oxygenase. , 1990, The Journal of biological chemistry.

[64]  A. Slusarenko,et al.  Arabidopsis is susceptible to infection by a downy mildew fungus. , 1990, The Plant cell.

[65]  F. Schuber,et al.  Mechanism of S-oxidation reactions catalyzed by a soybean hydroperoxide-dependent oxygenase , 1989 .

[66]  F. Durst,et al.  Hydroperoxide-dependent sulfoxidation catalyzed by soybean microsomes. , 1987, Archives of biochemistry and biophysics.

[67]  I. Rabino,et al.  Light, temperature, and anthocyanin production. , 1986, Plant physiology.

[68]  I. Yamazaki,et al.  Hydroperoxide-dependent hydroxylation involving "H2O2-reducible hemoprotein" in microsomes of pea seeds. A new type enzyme acting on hydroperoxide and a physiological role of seed lipoxygenase. , 1977, The Journal of biological chemistry.

[69]  D. Phillips,et al.  Lipoxygenase from potato tubers. Partial purification and properties of an enzyme that specifically oxygenates the 9-position of linoleic acid. , 1971, The Biochemical journal.

[70]  Z. Fan,et al.  Regulation of the flowering time of Arabidopsis thaliana by thylakoid ascorbate peroxidase , 2012 .

[71]  I. Mori,et al.  Biomarkers of green roof vegetation: anthocyanin and chlorophyll as stress marker pigments for plant stresses of roof environments , 2009 .

[72]  T. Aung,et al.  Stress-responsive alpha-dioxygenase expression in tomato roots. , 2005, Journal of Experimental Botany.

[73]  M. Hamberg,et al.  Involvement of the Arabidopsis alpha-DOX1 fatty acid dioxygenase in protection against oxidative stress and cell death. , 2002, The Plant journal : for cell and molecular biology.

[74]  J. Tzen,et al.  Cloning and secondary structure analysis of caleosin, a unique calcium-binding protein in oil bodies of plant seeds. , 1999, Plant & cell physiology.

[75]  T. Vernet,et al.  A family of yeast expression vectors containing the phage f1 intergenic region. , 1987, Gene.