ABSTRACT
Salicylic acid (SA) plays a central role in defense against pathogen attack, as well as in germination, flowering, senescence, and the acquisition of thermotolerance. In this report we investigate the involvement of phospholipase D (PLD) in the SA signaling pathway. In presence of exogenous primary alcohols, the production of phosphatidic acid by PLD is diverted toward the formation of phosphatidylalcohols through a reaction called transphosphatidylation. By in vivo metabolic phospholipid labeling with (33)P(i), PLD activity was found to be induced 45 min after addition of SA. We show that incubation of Arabidopsis (Arabidopsis thaliana) cell suspensions with primary alcohols inhibited the induction of two SA-responsive genes, PATHOGENESIS-RELATED1 and WRKY38, in a dose-dependent manner. This inhibitory effect was more pronounced when the primary alcohols were more hydrophobic. Secondary or tertiary alcohols had no inhibitory effect. These results provide compelling arguments for PLD activity being upstream of the induction of these genes by SA. A subsequent study of n-butanol effects on the SA-responsive transcriptome identified 1,327 genes differentially expressed upon SA treatment. Strikingly, the SA response of 380 of these genes was inhibited by n-butanol but not by tert-butanol. A detailed analysis of the regulation of these genes showed that PLD could act both positively and negatively, either on gene induction or gene repression. The overlap with the previously described phosphatidylinositol-4-kinase pathway is discussed.
Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/enzymology , Phospholipase D/metabolism , Salicylic Acid/metabolism , Signal Transduction , 1-Butanol/pharmacology , Arabidopsis/metabolism , Arabidopsis Proteins/chemistry , Arabidopsis Proteins/metabolism , Enzyme Activation , Gene Expression Profiling , Gene Expression Regulation, Plant , Oligonucleotide Array Sequence Analysis , Promoter Regions, Genetic , Transcription Factors/genetics , Transcription Factors/metabolism , tert-Butyl Alcohol/pharmacologyABSTRACT
Salicylic acid (SA) has a central role in defense against pathogen attack. In addition, its role in such diverse processes as germination, flowering, senescence, and thermotolerance acquisition has been documented. However, little is known about the early signaling events triggered by SA. Using Arabidopsis (Arabidopsis thaliana) suspension cells as a model, it was possible to show by in vivo metabolic phospholipid labeling with (33)P(i) that SA addition induced a rapid and early (in few minutes) decrease in a pool of phosphatidylinositol (PI). This decrease paralleled an increase in PI 4-phosphate and PI 4,5-bisphosphate. These changes could be inhibited by two different inhibitors of type III PI 4-kinases, phenylarsine oxide and 30 microm wortmannin; no inhibitory effect was seen with 1 microm wortmannin, a concentration inhibiting PI 3-kinases but not PI 4-kinases. We therefore undertook a study of the effects of wortmannin on SA-responsive transcriptomes. Using the Complete Arabidopsis Transcriptome MicroArray chip, we could identify 774 genes differentially expressed upon SA treatment. Strikingly, among these genes, the response to SA of 112 of them was inhibited by 30 microm wortmannin, but not by 1 microm wortmannin.
Subject(s)
1-Phosphatidylinositol 4-Kinase/metabolism , Arabidopsis/enzymology , Gene Expression Regulation, Plant , Phosphatidylinositols/metabolism , Salicylic Acid/metabolism , Androstadienes , Arabidopsis/genetics , Arabidopsis/metabolism , Cells, Cultured , Gene Expression Profiling , Phosphorus Radioisotopes , Promoter Regions, Genetic , WortmanninABSTRACT
The receptor for D-myo-inositol 1,4,5-trisphosphate (InsP3-R) has been well documented in animal cells. It constitutes an important component of the intracellular calcium signalling system. Today the corresponding genes in many species have been sequenced and the antibodies against some of the InsP3-Rs are available. In contrast, very little is known about its plant counterpart. Only a few published works have dealt directly with this topic. This review summarizes the available relevant data and determines some properties of putative plant receptor(s) including the in silico search for its gene in plant genomes, in vivo evidence, its electrophysiology, the parameters of InsP3-induced calcium release and InsP3 binding, immunological cross-reactivity, and subcellular localization. Future progress in this area seems to be inevitable as, despite the efforts, its gene in plants has not been identified yet.
