Separation of early and late responses to herbivory in Arabidopsis by changing plasmodesmal function.

Herbivory results in an array of physiological changes in the host that are separable from the associated physical damage. We have made the surprising observation that an Arabidopsis line (pdko3) mutated in genes encoding plasmodesmal proteins is defective in some, but not all, of the typical plant responses to herbivory. We tested the responses of plasma transmembrane potential (Vm) depolarization, voltage gated K(+) channel activity, cytosolic calcium [Ca2+]cyt and reactive oxygen species (ROS) (H2 O2 and NO) release, shoot-to-root signaling, biosynthesis of the phytohormone jasmonic acid (JA) and the elicitation of volatile organic compounds (VOCs). Following herbivory and the release of factors present in insect oral secretions (including a putative ß-galactofuranose polysaccharide), both the pdko3 and wild type (WT) plants showed a increased accumulation of [Ca2+]cyt , NO and H2 O2 . In contrast, unlike WT plants, the mutant line showed an almost complete loss of voltage gated K(+) channel activity and Vm depolarization, a loss of shoot-induced root-Vm depolarization, a loss of activation and regulation of gene expression of the JA defense pathway, and a much diminished release and altered profile of VOCs. The mutations in genes for plasmodesmal proteins have provided valuable genetic tools for the dissection of the complex spectrum of responses to herbivory and shown us that the responses to herbivory can be separated into a calcium-activated oxidative response and a K(+) -dependent Vm-activated jasmonate response associated with the release of VOCs.

Dynamics of membrane potential variation and gene expression induced by Spodoptera littoralis, Myzus persicae, and Pseudomonas syringae in Arabidopsis.

BACKGROUND: Biotic stress induced by various herbivores and pathogens invokes plant responses involving different defense mechanisms. However, we do not know whether different biotic stresses share a common response or which signaling pathways are involved in responses to different biotic stresses. We investigated the common and specific responses of Arabidopsis thaliana to three biotic stress agents: Spodoptera littoralis, Myzus persicae, and the pathogen Pseudomonas syringae. METHODOLOGY/PRINCIPAL FINDINGS: We used electrophysiology to determine the plasma membrane potential (V(m)) and we performed a gene microarray transcriptome analysis on Arabidopsis upon either herbivory or bacterial infection. V(m) depolarization was induced by insect attack; however, the response was much more rapid to S. littoralis (30 min -2 h) than to M. persicae (4-6 h). M. persicae differentially regulated almost 10-fold more genes than by S. littoralis with an opposite regulation. M. persicae modulated genes involved in flavonoid, fatty acid, hormone, drug transport and chitin metabolism. S. littoralis regulated responses to heat, transcription and ion transport. The latest Vm depolarization (16 h) was found for P. syringae. The pathogen regulated responses to salicylate, jasmonate and to microorganisms. Despite this late response, the number of genes differentially regulated by P. syringae was closer to those regulated by S. littoralis than by M. persicae. CONCLUSIONS/SIGNIFICANCE: Arabidopsis plasma membranes respond with a V(m) depolarization at times depending on the nature of biotic attack which allow setting a time point for comparative genome-wide analysis. A clear relationship between V(m) depolarization and gene expression was found. At V(m) depolarization timing, M. persicae regulates a wider array of Arabidopsis genes with a clear and distinct regulation than S. littoralis. An almost completely opposite regulation was observed between the aphid and the pathogen, with the former suppressing and the latter activating Arabidopsis defense responses.

Robotic mechanical wounding (MecWorm) versus herbivore-induced responses: early signaling and volatile emission in Lima bean (Phaseolus lunatus L.).

Insect herbivory on plants is a complex incident consisting of at least two different aspects, mechanical damage and chemical factors. Only the combination of both is able to induce the respective plant defenses. Thus, diverse plant species emit volatile organic compounds (VOCs) in response to herbivory (HW), whereas mechanical damage inflicted as single wounding event (MD) does not induce increased VOC emissions. In contrast, a robotic worm (MecWorm, MW) allowed demonstrating that continuous mechanical damage is sufficient to induce volatile emission in Lima bean. However, the induced VOC blends remain characteristic for the respective stimulus. In order to identify putative differences in plant signaling leading to defenses, we compared time courses of early signals induced by wounding in Lima bean. Neither MD nor MW alone was able to induce plasma membrane (V (m)) depolarization, as observed after Spodoptera littoralis HW, but V (m) depolarization occurred in both treatments when used in combination with herbivore-derived oral secretions. A significant increase in cytosolic Ca(2+) concentrations was observed only after HW, whereas MD and MW did not affect this second messenger. H(2)O(2) was generated within 2-3 h after leaf damage by HW and MW, whereas MD induced only half of the H(2)O(2) levels compared to the other treatments. Both HW and MW induced a marked accumulation of NO, but with distinct temporal patterns. NO production after MD followed the same trend but reached significantly lower values. The results indicate that chemical signals from the herbivores are responsible for the induction of the earliest signaling events. These changes appear to be characteristic for the reaction to herbivory.