In the absence of adaptive immunity displayed by animals, plants respond locally to biotic challenge via inducible basal defense networks activated through recognition and response to conserved pathogen-associated molecular patterns. In addition, immunity can be induced in tissues remote from infection sites by systemic acquired resistance (SAR), initiated after gene-for-gene recognition between plant resistance proteins and microbial effectors. The nature of the mobile signal and remotely activated networks responsible for establishing SAR remain unclear. Salicylic acid (SA) participates in the local and systemic response, but SAR does not require long-distance translocation of SA. Here, we show that, despite the absence of pathogen-associated molecular pattern contact, systemically responding leaves rapidly activate a SAR transcriptional signature with strong similarity to local basal defense. We present several lines of evidence that suggest jasmonates are central to systemic defense, possibly acting as the initiating signal for classic SAR. Jasmonic acid (JA), but not SA, rapidly accumulates in phloem exudates of leaves challenged with an avirulent strain of Pseudomonas syringae. In systemically responding leaves, transcripts associated with jasmonate biosynthesis are up-regulated within 4 h, and JA increases transiently. SAR can be mimicked by foliar JA application and is abrogated in mutants impaired in jasmonate synthesis or response. We conclude that jasmonate signaling appears to mediate long-distance information transmission. Moreover, the systemic transcriptional response shares extraordinary overlap with local herbivory and wounding responses, indicating that jasmonates may be pivotal to an evolutionarily conserved signaling network that decodes multiple abiotic and biotic stress signals.Arabidopsis thaliana ͉ jasmonic acid ͉ microarray ͉ Pseudomonas syringae

(N.S., S.B., M.J.M.)Lipases are involved in the generation of jasmonates, which regulate responses to biotic and abiotic stresses. Two sn-1-specific acyl hydrolases, DEFECTIVE IN ANTHER DEHISCENCE1 (DAD1) and DONGLE (DGL), have been reported to be localized in plastids and to be essential and sufficient for jasmonate biosynthesis in Arabidopsis (Arabidopsis thaliana) leaves. Here, we show that levels of 12-oxo-phytodienoic acid (OPDA) and jasmonic acid in three different DGL RNA interference lines and the dad1 mutant were similar to wild-type levels during the early wound response as well as after Pseudomonas infection. Due to the lack of sn-2 substrate specificity, synthesis of dinor OPDA was not expected and also not found to be affected in DGL knockdown and DGL-overexpressing lines. As reported, DAD1 participates in jasmonate formation only in the late wound response. In addition, DGL protein was found to be localized in lipid bodies and not in plastids. Furthermore, jasmonate levels in 16 additional mutants defective in the expression of lipases with predicted chloroplast localization did not show strong differences from wild-type levels after wounding, except for a phospholipase A (PLA) PLA-Ig1 (At1g06800) mutant line that displayed diminished wound-induced dinor OPDA, OPDA, and jasmonic acid levels. A quadruple mutant defective in four DAD1-like lipases displayed similar jasmonate levels as the mutant line of PLA-Ig1 after wounding. Hence, we identify PLAIg1 as a novel target gene to manipulate jasmonate biosynthesis. Our results suggest that, in addition to DAD1 and PLA-Ig1, still unidentified enzymes with sn-1 and sn-2 hydrolase activity are involved in wound-and pathogen-induced jasmonate formation, indicating functional redundancy within the lipase family.

Allene oxide synthase (AOS) is encoded by a single intronless gene in Arabidopsis thaliana (L.) Heynh. The promoter region of the AOS gene exhibits, in addition to the clements of a minimal promoter and the presence of general enhancers, cis-elements that, in other promoters, are responsible for stress- and ethyleneresponsiveness. Arabidopsis thaliana and Nicotiana tabacum L. were transformed with a chimaeric gene consisting of a 1.9-kb 5'-upstream sequence and the first 95 nucleotides of the AOS coding sequence translationally fused to uidA encoding beta-glucuronidase (GUS). Using histochemistry, GUS activity was seen in older leaves, in the bases of petioles and in stipules, during the early stages of carpel development, in maturing pollen grains and at the base of elongated filaments, as well as in abscission-zone scars. A role for jasmonates in floral organ abscission is suggested by these findings. Furthermore, the AOS promoter was activated both locally as well as systemically upon wounding. Jasmonic acid, 12-oxophytodienoic acid and coronatine strongly induced GUS activity. This induction remained confined to the treated leaf when agonists were applied locally to a leaf, suggesting that neither jasmonic acid nor 12-oxophytodienoic acid are physiologically relevant components of the systemic wound signal complex. Rather, the data show that jasmonates behave as local response regulators produced at or around the sites of action in response to appropriate triggers of their synthesis.

Allene oxide synthase (AOS) catalyzes the entrance reaction in the biosynthesis of the octadecanoids 12-oxophytodienoic acid (OPDA) and jasmonic acid (JA). The enzyme is feedback-regulated by JA and thus a target of the JA-signalling pathway. A fusion genetic approach was used to isolate mutants in this signalling pathway. Seeds from transgenic Arabidopsis thaliana plants expressing the Escherichia coli uidA gene encoding beta-glucuronidase (GUS) under the control of the AOS promoter were mutagenized with ethylmethane sulfonate and the progeny was screened for individuals exhibiting constitutive expression of uidA in the absence of an added octadecanoid. From 21,000 mutagenized plants, 8 lines showing constitutive AOS expression were obtained. The mutant lines were characterized further and fell into four classes, I to IV. All showed signs of growth inhibition encompassing both shoot and root systems, and accumulated higher than normal levels of OPDA. Mutants belonging to classes I and IV failed to set seeds due to defects in flower development which prevented self-pollination. One mutant, designated cas1, was characterized in more detail and showed, in addition to elevated levels of AOS mRNA, AOS polypeptide, OPDA, and JA, constitutive expression of JA-responsive genes ( VSP2, PDF1.2). The cas1 mutation is recessive and affects a single locus. Using cleaved amplified polymorphic sequences (CAPS) and simple sequence length polymorphisms (SSLP), the mutated gene was mapped to chromosome IV next to the SSLP marker CIW7.

A cyl hydrolases remodel biological membranes and release signaling molecules in response to a variety of biotic and abiotic stresses. After wounding or pathogen treatment lipases are necessary to release fatty acids as substrate for jasmonate biosynthesis. In osmotic stressed tissue they maintain integrity and functionality of membranes and during senescence lipases destroy and recycle membranes. Recently the role of several acyl hydrolases including DEFECTIVE IN ANTHER DEHISCENCE1 (DAD1) and DAD1-like lipase, e.g., DONGLE (DGL) and the phospholipase A (PLA) PLA-Iγ1 in jasmonate biosynthesis after wounding were investigated and functional redundancy within this family has been stated. Here we report necessity of diverse DAD1-like lipases in response to salt and sorbitol treatment. The lipase PLA-Iγ1 and PLA-Iβ2, which were both impaired in wound response, were also affected in response to osmotic stress in seed germination assays. Based on our observations and interpretations of transcription analyses generated by AtGenExpress project we speculate about more general roles of the DAD1-like lipase in diverse biological processes.