Identification of likely orthologs of tobacco salicylic acid-binding protein 2 and their role in systemic acquired resistance in Arabidopsis thaliana.

Salicylic acid-binding protein 2 (SABP2) is essential for the establishment of systemic acquired resistance (SAR) in tobacco; SABP2's methyl salicylate (MeSA) esterase activity is required in healthy systemic tissues of infected plants to release the active defense phytohormone SA from MeSA, which serves as a long-distance signal for SAR. In the current study, we characterize a new gene family from Arabidopsis thaliana encoding 18 potentially active alpha/beta fold hydrolases that share 32-57% identity with SABP2. Of 14 recombinant AtMES (MES for methyl esterase) proteins tested, five showed preference for MeSA as a substrate and displayed SA inhibition of MeSA esterase activity in vitro (AtMES1, -2, -4, -7, and -9). The two genes encoding MeSA esterases with the greatest activity, AtMES1 and -9, as well as AtMES7 were transcriptionally upregulated during infection of Arabidopsis with avirulent Pseudomonas syringae. In addition, conditional expression of AtMES1, -7, or -9 complemented SAR deficiency in SABP2-silenced tobacco, suggesting that these three members of the AtMES family are SABP2 functional homologs (orthologs). Underexpression by knockout mutation and/or RNAi-mediated silencing of multiple AtMES genes, including AtMES1, -2, -7, and -9, compromised SAR in Arabidopsis and correlated with enhanced accumulation of MeSA in the systemic tissue of SAR-induced plants. Together, the data show that several members of the AtMES gene family are functionally homologous to SABP2 and redundant for MeSA hydrolysis and probably SAR. These data suggest that MeSA is a conserved SAR signal in Arabidopsis and tobacco.

[1]  D. Klessig,et al.  Systemic acquired resistance: the elusive signal(s). , 2008, Current opinion in plant biology.

[2]  E. Pichersky,et al.  Inactive Methyl Indole-3-Acetic Acid Ester Can Be Hydrolyzed and Activated by Several Esterases Belonging to the AtMES Esterase Family of Arabidopsis1[W][OA] , 2008, Plant Physiology.

[3]  R. Welti,et al.  Plastid omega3-fatty acid desaturase-dependent accumulation of a systemic acquired resistance inducing activity in petiole exudates of Arabidopsis thaliana is independent of jasmonic acid. , 2007, The Plant journal : for cell and molecular biology.

[4]  Dhirendra Kumar,et al.  Methyl Salicylate Is a Critical Mobile Signal for Plant Systemic Acquired Resistance , 2007, Science.

[5]  Jonathan D. G. Jones,et al.  Pathological hormone imbalances. , 2007, Current opinion in plant biology.

[6]  J. Zeier,et al.  Pathogen-associated molecular pattern recognition rather than development of tissue necrosis contributes to bacterial induction of systemic acquired resistance in Arabidopsis. , 2007, The Plant journal : for cell and molecular biology.

[7]  M. Grant,et al.  Arabidopsis systemic immunity uses conserved defense signaling pathways and is mediated by jasmonates , 2007, Proceedings of the National Academy of Sciences.

[8]  S. Song,et al.  Overexpression of salicylic acid carboxyl methyltransferase reduces salicylic acid-mediated pathogen resistance in Arabidopsis thaliana , 2007, Plant Molecular Biology.

[9]  Dhirendra Kumar,et al.  Validation of RNAi silencing specificity using synthetic genes: salicylic acid-binding protein 2 is required for innate immunity in plants. , 2006, The Plant journal : for cell and molecular biology.

[10]  C. Wasternack,et al.  The Outcomes of Concentration-Specific Interactions between Salicylate and Jasmonate Signaling Include Synergy, Antagonism, and Oxidative Stress Leading to Cell Death , 2005, Plant Physiology.

[11]  J. Glazebrook Contrasting mechanisms of defense against biotrophic and necrotrophic pathogens. , 2005, Annual review of phytopathology.

[12]  F. Ausubel,et al.  Pseudomonas syringae manipulates systemic plant defenses against pathogens and herbivores. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Liang Tong,et al.  Structural and biochemical studies identify tobacco SABP2 as a methyl salicylate esterase and implicate it in plant innate immunity. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[14]  N. Buhot,et al.  Modulation of the biological activity of a tobacco LTP1 by lipid complexation. , 2004, Molecular biology of the cell.

[15]  M. Haring,et al.  Jasmonic Acid Is a Key Regulator of Spider Mite-Induced Volatile Terpenoid and Methyl Salicylate Emission in Tomato1[w] , 2004, Plant Physiology.

[16]  N. Yao,et al.  The role and regulation of programmed cell death in plant–pathogen interactions , 2004, Cellular microbiology.

[17]  R. Welti,et al.  The Arabidopsis thaliana Dihydroxyacetone Phosphate Reductase Gene SUPPRESSOR OF FATTY ACID DESATURASE DEFICIENCY1 Is Required for Glycerolipid Metabolism and for the Activation of Systemic Acquired Resistance On-line version contains Web-only data. , 2004, The Plant Cell Online.

[18]  Xinnian Dong,et al.  Systemic acquired resistance. , 2003, Annual review of phytopathology.

[19]  R. Hellens,et al.  pGreen: a versatile and flexible binary Ti vector for Agrobacterium-mediated plant transformation , 2000, Plant Molecular Biology.

[20]  Dhirendra Kumar,et al.  High-affinity salicylic acid-binding protein 2 is required for plant innate immunity and has salicylic acid-stimulated lipase activity , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Feng Chen,et al.  An Arabidopsis thaliana gene for methylsalicylate biosynthesis, identified by a biochemical genomics approach, has a role in defense. , 2003, The Plant journal : for cell and molecular biology.

[22]  Fumiaki Katagiri,et al.  Topology of the network integrating salicylate and jasmonate signal transduction derived from global expression phenotyping. , 2003, The Plant journal : for cell and molecular biology.

[23]  R. Dixon,et al.  A putative lipid transfer protein involved in systemic resistance signalling in Arabidopsis , 2002, Nature.

[24]  G. Martin,et al.  The tobacco salicylic acid-binding protein 3 (SABP3) is the chloroplast carbonic anhydrase, which exhibits antioxidant activity and plays a role in the hypersensitive defense response , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[25]  J. Dangl,et al.  A high-throughput method for quantifying growth of phytopathogenic bacteria in Arabidopsis thaliana. , 2002, The Plant journal : for cell and molecular biology.

[26]  P. Waterhouse,et al.  Construct design for efficient, effective and high-throughput gene silencing in plants. , 2001, The Plant journal : for cell and molecular biology.

[27]  N. Chua,et al.  Technical advance: An estrogen receptor-based transactivator XVE mediates highly inducible gene expression in transgenic plants. , 2000, The Plant journal : for cell and molecular biology.

[28]  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.

[29]  J. Parker,et al.  Different requirements for EDS1 and NDR1 by disease resistance genes define at least two R gene-mediated signaling pathways in Arabidopsis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[30]  I. Raskin,et al.  Endogenous Methyl Salicylate in Pathogen-Inoculated Tobacco Plants , 1998 .

[31]  Ilya Raskin,et al.  Airborne signalling by methyl salicylate in plant pathogen resistance , 1997, Nature.

[32]  D. Klessig,et al.  Identification of a salicylic acid-responsive element in the promoter of the tobacco pathogenesis-related beta-1,3-glucanase gene, PR-2d. , 1996, The Plant journal : for cell and molecular biology.

[33]  E. Ward,et al.  Salicylic Acid Is Not the Translocated Signal Responsible for Inducing Systemic Acquired Resistance but Is Required in Signal Transduction. , 1994, The Plant cell.

[34]  R. Dixon,et al.  Biologically induced systemic acquired resistance in Arabidopsis thaliana , 1994 .

[35]  F. Corpet Multiple sequence alignment with hierarchical clustering. , 1988, Nucleic acids research.

[36]  King Eo,et al.  Two simple media for the demonstration of pyocyanin and fluorescin. , 1954 .

[37]  E. King,et al.  Two simple media for the demonstration of pyocyanin and fluorescin. , 1954, The Journal of laboratory and clinical medicine.