A novel cold-inducible gene from Arabidopsis, RCI3, encodes a peroxidase that constitutes a component for stress tolerance.

A cDNA from Arabidopsis corresponding to a new cold-inducible gene, RCI3 (for Rare Cold Inducible gene 3), was isolated. Isoelectric focusing electrophoresis and staining of peroxidase activity demonstrated that RCI3 encodes an active cationic peroxidase. RNA-blot analysis revealed that RCI3 expression in response to low temperature is negatively regulated by light, as RCI3 transcripts were exclusively detected in etiolated seedlings and roots of adult plants. RCI3 expression was also induced in etiolated seedlings, but not in roots, exposed to dehydration, salt stress or ABA, indicating that it is subjected to a complex regulation through different signaling pathways. Analysis of transgenic plants containing RCI3::GUS fusions established that this regulation occurs at the transcriptional level during plant development, and that cold-induced RCI3 expression in roots is mainly restricted to the endodermis. Plants overexpressing RCI3 showed an increase in dehydration and salt tolerance, while antisense suppression of RCI3 expression gave dehydration- and salt-sensitive phenotypes. These results indicate that RCI3 is involved in the tolerance to both stresses in Arabidopsis, and illustrate that manipulation of RCI3 has a potential with regard to plant improvement of stress tolerance.

[1]  R. Catalá,et al.  Developmental and stress regulation of RCI2A and RCI2B, two cold-inducible genes of arabidopsis encoding highly conserved hydrophobic proteins. , 2001, Plant physiology.

[2]  H. Greppin,et al.  Molecular cloning and tissue-specific expression of an anionic peroxidase in zucchini. , 1999, Plant physiology.

[3]  N. Raikhel,et al.  Intracellular trafficking of secretory proteins , 1992, Plant Molecular Biology.

[4]  H. Jespersen,et al.  Sequence and RT-PCR expression analysis of two peroxidases from Arabidopsis thaliana belonging to a novel evolutionary branch of plant peroxidases , 1997, Plant Molecular Biology.

[5]  S. H. Lee,et al.  A novel cold-inducible zinc finger protein from soybean, SCOF-1, enhances cold tolerance in transgenic plants. , 2001, The Plant journal : for cell and molecular biology.

[6]  F. Madueño,et al.  Dimerization of Arabidopsis 14‐3‐3 proteins: structural requirements within the N‐terminal domain and effect of calcium , 1999, FEBS letters.

[7]  J. Terol,et al.  The Arabidopsis CBF gene family is composed of three genes encoding AP2 domain-containing proteins whose expression Is regulated by low temperature but not by abscisic acid or dehydration. , 1999, Plant physiology.

[8]  D. Siminovitch,et al.  Correlation between Cold- and Drought-Induced Frost Hardiness in Winter Wheat and Rye Varieties. , 1982, Plant physiology.

[9]  F. Sarhan,et al.  A leaf-specific gene stimulated by light during wheat acclimation to low temperature , 1993, Plant Molecular Biology.

[10]  E. T. Palva,et al.  Differential expression of two related, low-temperature-induced genes in Arabidopsis thaliana (L.) Heynh , 1993, Plant Molecular Biology.

[11]  F. Skoog,et al.  A revised medium for rapid growth and bio assays with tobacco tissue cultures , 1962 .

[12]  J. Braam,et al.  Arabidopsis TCH3 encodes a novel Ca2+ binding protein and shows environmentally induced and tissue-specific regulation. , 1994, The Plant cell.

[13]  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.

[14]  Kazuo Shinozaki,et al.  Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor , 1999, Nature Biotechnology.

[15]  H. Okada,et al.  Structure of the horseradish peroxidase isozyme C genes. , 1988, European journal of biochemistry.

[16]  K. Shinozaki,et al.  Two Transcription Factors, DREB1 and DREB2, with an EREBP/AP2 DNA Binding Domain Separate Two Cellular Signal Transduction Pathways in Drought- and Low-Temperature-Responsive Gene Expression, Respectively, in Arabidopsis , 1998, Plant Cell.

[17]  K. Shinozaki,et al.  Arabidopsis DNA Encoding Two Desiccation-Responsive rd29 Genes , 1993, Plant physiology.

[18]  E. Nilsen,et al.  Physiology of Plants Under Stress , 1987 .

[19]  K. Shinozaki,et al.  A novel cis-acting element in an Arabidopsis gene is involved in responsiveness to drought, low-temperature, or high-salt stress. , 1994, The Plant cell.

[20]  E. Stockinger,et al.  Arabidopsis thaliana CBF1 encodes an AP2 domain-containing transcriptional activator that binds to the C-repeat/DRE, a cis-acting DNA regulatory element that stimulates transcription in response to low temperature and water deficit. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[21]  A. Leyva,et al.  Low Temperature Induces the Accumulation of Phenylalanine Ammonia-Lyase and Chalcone Synthase mRNAs of Arabidopsis thaliana in a Light-Dependent Manner , 1995, Plant physiology.

[22]  D. Bowles,et al.  Defense-related proteins in higher plants. , 1990, Annual review of biochemistry.

[23]  S. J. Gilmour,et al.  Constitutive expression of the cold-regulated Arabidopsis thaliana COR15a gene affects both chloroplast and protoplast freezing tolerance. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[24]  P. S. Kim,et al.  Evidence that the leucine zipper is a coiled coil. , 1989, Science.

[25]  Julio Salinas,et al.  Two related low-temperature-inducible genes of Arabidopsis encode proteins showing high homology to 14-3-3 proteins, a family of putative kinase regulators , 1994, Plant Molecular Biology.

[26]  Marc Van Montagu,et al.  Efficient octopine Ti plasmid-derived vectors for Agrobacterium- mediated gene transfer to plants , 1985, Nucleic Acids Res..

[27]  Michael F. Thomashow,et al.  PLANT COLD ACCLIMATION: Freezing Tolerance Genes and Regulatory Mechanisms. , 1999, Annual review of plant physiology and plant molecular biology.

[28]  P. Harrison,et al.  Localization of expression of three cold-induced genes, blt101, blt4. 9, and blt14, in different tissues of the crown and developing leaves of cold-acclimated cultivated barley , 1998, Plant physiology.

[29]  R. Bressan,et al.  Characterization and in situ localization of a salt-induced tomato peroxidase mRNA , 1994, Plant Molecular Biology.

[30]  R. T. Cruz,et al.  Structural Changes and Associated Reduction of Hydraulic Conductance in Roots of Sorghum bicolor L. following Exposure to Water Deficit. , 1992, Plant physiology.

[31]  V. Valpuesta,et al.  Expression of a highly basic peroxidase gene in NaCl‐adapted tomato cell suspensions , 1997, FEBS letters.

[32]  L. Willmitzer,et al.  Improved method for the isolation of RNA from plant tissues. , 1987, Analytical biochemistry.

[33]  D. Baulcombe,et al.  Expression of biologically active viral satellite RNA from the nuclear genome of transformed plants , 1986, Nature.

[34]  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.

[35]  O. Schabenberger,et al.  Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance. , 1998, Science.

[36]  K. Shinozaki,et al.  Molecular responses to dehydration and low temperature: differences and cross-talk between two stress signaling pathways. , 2000, Current opinion in plant biology.

[37]  E. Steudle.  Review article. How does water get through roots , 1998 .

[38]  Ylva Gavel,et al.  Sequence differences between glycosylated and non-glycosylated Asn-X-Thr/Ser acceptor sites: implications for protein engineering , 1990, Protein engineering.

[39]  M. Delseny,et al.  The Brassica oleracea rDNA spacer revisited , 1992, Plant Molecular Biology.

[40]  J. Brown,et al.  Arabidopsis intron mutations and pre-mRNA splicing. , 1996, The Plant journal : for cell and molecular biology.

[41]  J. Levitt,et al.  Responses of Plants to Environmental Stress, 2nd Edition, Volume 1: Chilling, Freezing, and High Temperature Stresses. , 1980 .

[42]  S. Rasmussen,et al.  cDNA, amino acid and carbohydrate sequence of barley seed-specific peroxidase BP 1 , 1992, Plant Molecular Biology.

[43]  M. Tester,et al.  Cell-type-specific calcium responses to drought, salt and cold in the Arabidopsis root. , 2000, The Plant journal : for cell and molecular biology.

[44]  K. Welinder.  Superfamily of plant, fungal and bacterial peroxidases , 1992 .

[45]  L. Gusta,et al.  Abscisic Acid-induced freezing resistance in cultured plant cells. , 1983, Plant physiology.

[46]  V. Valpuesta,et al.  Induction of a tomato peroxidase gene in vascular tissue , 1994, FEBS letters.

[47]  E. Steudle,et al.  How does water get through roots , 1998 .

[48]  S. Fry,et al.  Arabidopsis TCH4, regulated by hormones and the environment, encodes a xyloglucan endotransglycosylase. , 1995, The Plant cell.

[49]  J. A. Jarillo,et al.  Two Homologous Low-Temperature-Inducible Genes from Arabidopsis Encode Highly Hydrophobic Proteins , 1997, Plant physiology.

[50]  E. Stockinger,et al.  Low temperature regulation of the Arabidopsis CBF family of AP2 transcriptional activators as an early step in cold-induced COR gene expression. , 1998, The Plant journal : for cell and molecular biology.

[51]  E. T. Palva,et al.  Role of Abscisic Acid in Drought-Induced Freezing Tolerance, Cold Acclimation, and Accumulation of LT178 and RAB18 Proteins in Arabidopsis thaliana , 1995, Plant physiology.

[52]  P. Hasegawa,et al.  Nucleotide Sequences of Two Peroxidase Genes from Tomato (Lycopersicon esculentum) , 1993, Plant physiology.

[53]  S. Tähtiharju,et al.  Antisense inhibition of protein phosphatase 2C accelerates cold acclimation in Arabidopsis thaliana. , 2001, The Plant journal : for cell and molecular biology.

[54]  C. Somerville,et al.  Sulfonylurea-resistant mutants of Arabidopsis thaliana , 1986, Molecular and General Genetics MGG.