Salicylic acid dependent signaling promotes basal thermotolerance but is not essential for acquired thermotolerance in Arabidopsis thaliana.

Salicylic acid (SA) is reported to protect plants from heat shock (HS), but insufficient is known about its role in thermotolerance or how this relates to SA signaling in pathogen resistance. We tested thermotolerance and expression of pathogenesis-related (PR) and HS proteins (HSPs) in Arabidopsis thaliana genotypes with modified SA signaling: plants with the SA hydroxylase NahG transgene, the nonexpresser of PR proteins (npr1) mutant, and the constitutive expressers of PR proteins (cpr1 and cpr5) mutants. At all growth stages from seeds to 3-week-old plants, we found evidence for SA-dependent signaling in basal thermotolerance (i.e. tolerance of HS without prior heat acclimation). Endogenous SA correlated with basal thermotolerance, with the SA-deficient NahG and SA-accumulating cpr5 genotypes having lowest and highest thermotolerance, respectively. SA promoted thermotolerance during the HS itself and subsequent recovery. Recovery from HS apparently involved an NPR1-dependent pathway but thermotolerance during HS did not. SA reduced electrolyte leakage, indicating that it induced membrane thermoprotection. PR-1 and Hsp17.6 were induced by SA or HS, indicating common factors in pathogen and HS responses. SA-induced Hsp17.6 expression had a different dose-response to PR-1 expression. HS-induced Hsp17.6 protein appeared more slowly in NahG. However, SA only partially induced HSPs. Hsp17.6 induction by HS was more substantial than by SA, and we found no SA effect on Hsp101 expression. All genotypes, including NahG and npr1, were capable of expression of HSPs and acquisition of HS tolerance by prior heat acclimation. Although SA promotes basal thermotolerance, it is not essential for acquired thermotolerance.

[1]  J. Draper,et al.  Compromising early salicylic acid accumulation delays the hypersensitive response and increases viral dispersal during lesion establishment in TMV-infected tobacco. , 1997, The Plant journal : for cell and molecular biology.

[2]  J. Dean,et al.  Formation and vacuolar localization of salicylic acid glucose conjugates in soybean cell suspension cultures , 2003 .

[3]  E. Vierling,et al.  Synthesis of Small Heat-Shock Proteins Is Part of the Developmental Program of Late Seed Maturation , 1996, Plant physiology.

[4]  H. Goodman,et al.  DNA sequence analysis of a PR-1a gene from tobacco: Molecular relationship of heat shock and pathogen responses in plants , 1988, Molecular and General Genetics MGG.

[5]  E. Richards,et al.  Induced instability of two Arabidopsis constitutive pathogen-response alleles , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[6]  E. Vierling,et al.  Mutants of Arabidopsis thaliana defective in the acquisition of tolerance to high temperature stress. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[7]  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.

[8]  J. Dat,et al.  Hydrogen peroxide‐ and glutathione‐associated mechanisms of acclimatory stress tolerance and signalling , 1997 .

[9]  J. Draper Plant Genetic Transformation and Gene Expression: A Laboratory Manual , 1988 .

[10]  D F Klessig,et al.  The cpr5 mutant of Arabidopsis expresses both NPR1-dependent and NPR1-independent resistance. , 1997, The Plant cell.

[11]  K. Morris,et al.  Salicylic acid has a role in regulating gene expression during leaf senescence. , 2000, The Plant journal : for cell and molecular biology.

[12]  M. Knight,et al.  Protection against Heat Stress-Induced Oxidative Damage in Arabidopsis Involves Calcium, Abscisic Acid, Ethylene, and Salicylic Acid , 2002, Plant Physiology.

[13]  M. Matsuoka,et al.  Synthesis of Stress Proteins in Tobacco Leaves , 1985 .

[14]  Lin Qiu,et al.  Salicylate Triggers Heat Shock Factor Differently than Heat (*) , 1995, The Journal of Biological Chemistry.

[15]  K. Dixon,et al.  Acetyl salicylic acid (Aspirin) and salicylic acid induce multiple stress tolerance in bean and tomato plants , 2000, Plant Growth Regulation.

[16]  J. L. Key,et al.  Solute Leakage in Soybean Seedlings under Various Heat Shock Regimes , 1985 .

[17]  S. Rüdiger,et al.  Identification of thermolabile Escherichia coli proteins: prevention and reversion of aggregation by DnaK and ClpB , 1999, The EMBO journal.

[18]  S. Seo,et al.  Induction of Salicylic Acid β-Glucosidase in Tobacco Leaves by Exogenous Salicylic Acid , 1995 .

[19]  J. Lee,et al.  Derepression of the activity of genetically engineered heat shock factor causes constitutive synthesis of heat shock proteins and increased thermotolerance in transgenic Arabidopsis. , 1995, The Plant journal : for cell and molecular biology.

[20]  C. Wang,et al.  Reduction of chilling injury and transcript accumulation of heat shock proteins in tomato fruit by methyl jasmonate and methyl salicylate , 2001 .

[21]  C. Pollock,et al.  Development of laboratory-based methods for assessing seedling thermotolerance in pearl millet , 1997 .

[22]  I. Graham,et al.  Long-chain acyl-CoA oxidases of Arabidopsis. , 1999, The Plant journal : for cell and molecular biology.

[23]  J. Draper,et al.  Characterisation of a wound-induced transcript from the monocot asparagus that shares similarity with a class of intracellular pathogenesis-related (PR) proteins , 1992, Plant Molecular Biology.

[24]  J. Glazebrook,et al.  Loss of non-host resistance of Arabidopsis NahG to Pseudomonas syringae pv. phaseolicola is due to degradation products of salicylic acid. , 2003, The Plant journal : for cell and molecular biology.

[25]  S. Potter,et al.  Regulation of pathogenesis-related protein-1a gene expression in tobacco. , 1993, The Plant cell.

[26]  E. Vierling,et al.  Hsp101 is necessary for heat tolerance but dispensable for development and germination in the absence of stress. , 2001, The Plant journal : for cell and molecular biology.

[27]  M. Cronjé,et al.  Salicylic acid influences Hsp70/Hsc70 expression in Lycopersicon esculentum: dose- and time-dependent induction or potentiation. , 1999, Biochemical and biophysical research communications.

[28]  K B Singh,et al.  The auxin, hydrogen peroxide and salicylic acid induced expression of the Arabidopsis GST6 promoter is mediated in part by an ocs element. , 1999, The Plant journal : for cell and molecular biology.

[29]  J. Dat,et al.  Effects of Salicylic Acid on Oxidative Stress and Thermotolerance in Tobacco , 2000 .

[30]  Xinnian Dong,et al.  Nuclear Localization of NPR1 Is Required for Activation of PR Gene Expression , 2000, Plant Cell.

[31]  Malik,et al.  Modified expression of a carrot small heat shock protein gene, hsp17. 7, results in increased or decreased thermotolerancedouble dagger , 1999, The Plant journal : for cell and molecular biology.

[32]  E. Vierling,et al.  Arabidopsis hot Mutants Define Multiple Functions Required for Acclimation to High Temperatures1 , 2003, Plant Physiology.

[33]  F. Ausubel,et al.  Roles of Salicylic Acid, Jasmonic Acid, and Ethylene in cpr-Induced Resistance in Arabidopsis , 2000, Plant Cell.

[34]  J. Dat,et al.  Changes in salicylic acid and antioxidants during induced thermotolerance in mustard seedlings , 1998, Plant physiology.

[35]  Xinnian Dong,et al.  Characterization of an Arabidopsis Mutant That Is Nonresponsive to Inducers of Systemic Acquired Resistance. , 1994, The Plant cell.

[36]  G. Ponce,et al.  Maize HSP101 Plays Important Roles in Both Induced and Basal Thermotolerance and Primary Root Growth Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.010487. , 2002, The Plant Cell Online.

[37]  J. Görlach,et al.  Growth Stage–Based Phenotypic Analysis of Arabidopsis , 2001, The Plant Cell Online.

[38]  J. Draper,et al.  Isolation of an asparagus intracellular PR gene (AoPR1) wound-responsive promoter by the inverse polymerase chain reaction and its characterization in transgenic tobacco. , 1993, The Plant journal : for cell and molecular biology.

[39]  M. Van Montagu,et al.  At-HSP17.6A, encoding a small heat-shock protein in Arabidopsis, can enhance osmotolerance upon overexpression. , 2001, The Plant journal : for cell and molecular biology.

[40]  E. Ward,et al.  A Central Role of Salicylic Acid in Plant Disease Resistance , 1994, Science.

[41]  M. Tal,et al.  Effects of Dehydration and High Temperature on the Stability of Leaf Membranes of Lycoperskon esculentum, L. cheesmanii, L. peruvianum and Solanum pennellii , 1983 .

[42]  K. Mendgen,et al.  PR-1 protein inhibits the differentiation of rust infection hyphae in leaves of acquired resistant broad bean. , 1999, The Plant journal : for cell and molecular biology.

[43]  G. Balogh,et al.  Synechocystis HSP17 is an amphitropic protein that stabilizes heat-stressed membranes and binds denatured proteins for subsequent chaperone-mediated refolding , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[44]  S. Lindquist,et al.  Heat Shock Protein 101 Plays a Crucial Role in Thermotolerance in Arabidopsis , 2000, Plant Cell.

[45]  K. Hinderhofer,et al.  HSF3, a new heat shock factor from Arabidopsis thaliana, derepresses the heat shock response and confers thermotolerance when overexpressed in transgenic plants , 1998, Molecular and General Genetics MGG.

[46]  G. Pastori,et al.  Common Components, Networks, and Pathways of Cross-Tolerance to Stress. The Central Role of “Redox” and Abscisic Acid-Mediated Controls1 , 2002, Plant Physiology.

[47]  L. C. Loon,et al.  The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins , 1999 .

[48]  D. Klessig,et al.  A mutation in Arabidopsis that leads to constitutive expression of systemic acquired resistance. , 1994, The Plant cell.

[49]  J. H. Williams,et al.  Temperature Tolerance in Soybeans. I. Evaluation of a Technique for Assessing Cellular Membrane Thermostability 1 , 1979 .

[50]  B. Kunkel,et al.  Cross talk between signaling pathways in pathogen defense. , 2002, Current opinion in plant biology.

[51]  D. Klessig,et al.  Temperature-Dependent Induction of Salicylic Acid and Its Conjugates during the Resistance Response to Tobacco Mosaic Virus Infection. , 1992, The Plant cell.

[52]  Akira Watanabe,et al.  Identification of a novel gene HYS1/CPR5 that has a repressive role in the induction of leaf senescence and pathogen-defence responses in Arabidopsis thaliana. , 2002, The Plant journal : for cell and molecular biology.

[53]  W. Gurley,et al.  HSP101: A Key Component for the Acquisition of Thermotolerance in Plants , 2000, Plant Cell.

[54]  S. Potter,et al.  Acquired resistance in Arabidopsis. , 1992, The Plant cell.

[55]  A. Srinivasan,et al.  Heat tolerance in food legumes as evaluated by cell membrane thermostability and chlorophyll fluorescence techniques , 2004, Euphytica.

[56]  M. Margis-Pinheiro,et al.  Bean class IV chitinase gene: structure, developmental expression and induction by heat stress , 1994 .

[57]  Zheng-Hua Ye,et al.  Mutation of a Chitinase-Like Gene Causes Ectopic Deposition of Lignin, Aberrant Cell Shapes, and Overproduction of Ethylene Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.010278. , 2002, The Plant Cell Online.

[58]  I. Horváth,et al.  Membrane lipid perturbation modifies the set point of the temperature of heat shock response in yeast. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[59]  J. Arias,et al.  Auxin-induced Stress Potentiates trans-activation by a Conserved Plant Basic/Leucine-zipper Factor* , 1998, The Journal of Biological Chemistry.

[60]  J. Dat,et al.  Parallel changes in H2O2 and catalase during thermotolerance induced by salicylic acid or heat acclimation in mustard seedlings , 1998, Plant physiology.

[61]  Induction of thermotolerance in potato microplants by acetylsalicylic acid and H2O2 , 1998 .

[62]  E. W. Hamilton,et al.  Heat-shock proteins are induced in unstressed leaves of Nicotiana attenuata (Solanaceae) when distant leaves are stressed. , 2001, American journal of botany.

[63]  Xinnian Dong,et al.  In Vivo Interaction between NPR1 and Transcription Factor TGA2 Leads to Salicylic Acid–Mediated Gene Activation in Arabidopsis Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.001628. , 2002, The Plant Cell Online.

[64]  Enwu Liu,et al.  The Arabidopsis NPR1/NIM1 Protein Enhances the DNA Binding Activity of a Subgroup of the TGA Family of bZIP Transcription Factors , 2000, Plant Cell.

[65]  F. Schöffl,et al.  Developmental regulation and tissue-specific differences of heat shock gene expression in transgenic tobacco and Arabidopsis plants , 1995, Plant Molecular Biology.

[66]  E. Lam,et al.  NPR1 differentially interacts with members of the TGA/OBF family of transcription factors that bind an element of the PR-1 gene required for induction by salicylic acid. , 2000, Molecular plant-microbe interactions : MPMI.