Elodie Vandelle

Constitutive cyclic GMP accumulation in Arabidopsis thaliana compromises systemic acquired resistance induced by an avirulent pathogen by modulating local signals

The infection of Arabidopsis thaliana plants with avirulent pathogens causes the accumulation of cGMP with a biphasic profile downstream of nitric oxide signalling. However, plant enzymes that modulate cGMP levels have yet to be identified, so we generated transgenic A. thaliana plants expressing the rat soluble guanylate cyclase (GC) to increase genetically the level of cGMP and to study the function of cGMP in plant defence responses. Once confirmed that cGMP levels were higher in the GC transgenic lines than in wild-type controls, the GC transgenic plants were then challenged with bacterial pathogens and their defence responses were characterized. Although local resistance was similar in the GC transgenic and wild-type lines, differences in the redox state suggested potential cross-talk between cGMP and the glutathione redox system. Furthermore, large-scale transcriptomic and proteomic analysis highlighted the significant modulation of both gene expression and protein abundance at the infection site, inhibiting the establishment of systemic acquired resistance. Our data indicate that cGMP plays a key role in local responses controlling the induction of systemic acquired resistance in plants challenged with avirulent pathogens.

S-nitrosylation of peroxiredoxin II E promotes peroxynitrite-mediated tyrosine nitration

Nitric oxide (NO) is a free radical product of cell metabolism that plays diverse and important roles in the regulation of cellular function. S-Nitrosylation is emerging as a specific and fundamental posttranslational protein modification for the transduction of NO bioactivity, but very little is known about its physiological functions in plants. We investigated the molecular mechanism for S-nitrosylation of peroxiredoxin II E (PrxII E) from Arabidopsis thaliana and found that this posttranslational modification inhibits the hydroperoxide-reducing peroxidase activity of PrxII E, thus revealing a novel regulatory mechanism for peroxiredoxins. Furthermore, we obtained biochemical and genetic evidence that PrxII E functions in detoxifying peroxynitrite (ONOO−), a potent oxidizing and nitrating species formed in a diffusion-limited reaction between NO and O2− that can interfere with Tyr kinase signaling through the nitration of Tyr residues. S-Nitrosylation also inhibits the ONOO− detoxification activity of PrxII E, causing a dramatic increase of ONOO−-dependent nitrotyrosine residue formation. The same increase was observed in a prxII E mutant line after exposure to ONOO−, indicating that the PrxII E modulation of ONOO− bioactivity is biologically relevant. We conclude that NO regulates the effects of its own radicals through the S-nitrosylation of crucial components of the antioxidant defense system that function as common triggers for reactive oxygen species– and NO-mediated signaling events.

NO signals in the haze: Nitric oxide signalling in plant defence

Nitric oxide (NO) is gaining increasing attention as a regulator of diverse (patho-)physiological processes in plants. Although this molecule has been described as playing a role in numerous conditions, its production, turnover and mode of action are poorly understood. Recent studies on NO production have tended to highlight the questions that still remain unanswered rather than telling us more about NO metabolism. But regarding NO signalling and functions, new findings have given an impression of the intricacy of NO-related signalling networks. Different targets of protein S-nitrosylation have been characterised and enzymatic routes controlling this posttranslational modification are emerging, along with their physiological implications. Evidence is also accumulating for protein tyrosine nitration and cGMP as important components of NO-related signal transduction.

Protein nitration during defense response in Arabidopsis thaliana

Nitric oxide and reactive oxygen species play a key role in the plant hypersensitive disease resistance response, and protein tyrosine nitration is emerging as an important mechanism of their co‐operative interaction. Up to now, the proteins targeted by this post‐translational modification in plants are still totally unknown. In this study, we analyzed for the first time proteins undergoing nitration during the hypersensitive response by analyzing via 1D‐ and 2D‐western blot the protein extracts from Arabidopsis thaliana plants challenged with an avirulent bacterial pathogen (Pseudomonas syringae pv. Tomato). We show that the plant disease resistance response is correlated with a modulation of nitration of proteins involved in important cellular process, such as photosynthesis, glycolysis and nitrate assimilation. These findings shed new light on the signaling functions of nitric oxide and reactive oxygen species, paving the way on studies on the role of this post‐translational modification in plants.

Peroxynitrite formation and function in plants

Peroxynitrite (ONOO(-)) is a reactive nitrogen species formed when nitric oxide (NO) reacts with the superoxide anion (O(2)(-)). It was first identified as a mediator of cell death in animals but was later shown to act as a positive regulator of cell signaling, mainly through the posttranslational modification of proteins by tyrosine nitration. In plants, peroxynitrite is not involved in NO-mediated cell death and its physiological function is poorly understood. However, it is emerging as a potential signaling molecule during the induction of defense responses against pathogens and this could be mediated by the selective nitration of tyrosine residues in a small number of proteins. In this review we discuss the general role of tyrosine nitration in plants and evaluate recent evidence suggesting that peroxynitrite is an effector of NO-mediated signaling following pathogen infection.

Detection and function of nitric oxide during the hypersensitive response in Arabidopsis thaliana: where there's a will there's a way.

Nitric oxide (NO) was identified as a key player in plant defence responses approximately 20 years ago and a large body of evidence has accumulated since then supporting its role as a signalling molecule. However, there are many discrepancies in current NO detection assays and the enzymatic pathways responsible for its synthesis have yet to be determined. This has provoked strong debates concerning the function of NO in plants, even questioning its existence in planta. Here we gather data obtained using the model pathosystem Arabidopsis/Pseudomonas, which confirms the production of NO during the hypersensitive response and supports is role as a trigger of hypersensitive cell death and a mediator of defence gene expression. Finally, we discuss potential sources of NO synthesis, focusing on the role of nitrite as major substrate for NO production during incompatible interactions.

Methods for nitric oxide detection during plant-pathogen interactions.

Nitric oxide (NO) is involved in the transduction of numerous signals in living organisms, and its biological effects are often influenced by its concentration. Therefore, the ability to reliably detect and quantify NO is crucial to understanding its role in cellular processes. Many techniques are available to detect and quantify NO, but depending on the material and the aim of the analysis, specific adaptations are often required because its high chemical reactivity leads to the formation of numerous reactive nitrogen species that make the accurate determination of NO levels difficult. Moreover, the pathogen-induced hypersensitive response leads to high rates of reactive oxygen species production that react with NO and lead to the formation of its oxidized derivates. The aim of this chapter is to provide an overview of the methods that have so far been employed to detect and measure NO in plants during the hypersensitive disease resistance response.

Genome-wide characterisation and expression profile of the grapevine ATL ubiquitin ligase family reveal biotic and abiotic stress-responsive and development-related members

The Arabidopsis Tóxicos en Levadura (ATL) protein family is a class of E3 ubiquitin ligases with a characteristic RING-H2 Zn-finger structure that mediates diverse physiological processes and stress responses in plants. We carried out a genome-wide survey of grapevine (Vitis vinifera L.) ATL genes and retrieved 96 sequences containing the canonical ATL RING-H2 domain. We analysed their genomic organisation, gene structure and evolution, protein domains and phylogenetic relationships. Clustering revealed several clades, as already reported in Arabidopsis thaliana and rice (Oryza sativa), with an expanded subgroup of grapevine-specific genes. Most of the grapevine ATL genes lacked introns and were scattered among the 19 chromosomes, with a high level of duplication retention. Expression profiling revealed that some ATL genes are expressed specifically during early or late development and may participate in the juvenile to mature plant transition, whereas others may play a role in pathogen and/or abiotic stress responses, making them key candidates for further functional analysis. Our data offer the first genome-wide overview and annotation of the grapevine ATL family, and provide a basis for investigating the roles of specific family members in grapevine physiology and stress responses, as well as potential biotechnological applications.

Nitric Oxide-Mediated Signaling Functions During the Plant Hypersensitive Response

Growing evidence suggests that nitric oxide (NO) is a central molecule in several physiological functions, ranging from plant development to defence responses. Plants use NO as a signaling molecule in pathways comparable to those of mammals, suggesting the existence of many commonalities between the action of NO in plants and animals.

Host-mediated S-nitrosylation disarms the bacterial effector HopAI1 to re-establish immunity

S-nitrosylation of the bacterial effector HopAI1 during plant infection is a nitric oxide-dependent host strategy to disarm the effector activity and reestablish immunity Pathogens deliver effectors into plant cells to suppress immunity-related signaling. However, effector recognition by the host elicits a hypersensitive response (HR) that overcomes the inhibition of host signaling networks, restoring disease resistance. Signaling components are shared between the pathogen-associated molecular pattern-triggered immunity and effector-triggered immunity, and it is unclear how plants inactivate these effectors to execute the HR. Here, we report that, in Arabidopsis thaliana, during the onset of the HR, the bacterial effector HopAI1 is S-nitrosylated and that this modification inhibits its phosphothreonine lyase activity. HopAI1 targets and suppresses mitogen-activated protein kinases (MAPKs). The S-nitrosylation of HopAI1 restores MAPK signaling and is required during the HR for activation of the associated cell death. S-nitrosylation is therefore revealed here as a nitric oxide-dependent host strategy involved in plant immunity that works by directly disarming effector proteins.