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Localisation and dynamics of free and bound 12-oxo-phytodienoic acid (OPDA) pools in Arabidopsis

English title Localisation and dynamics of free and bound 12-oxo-phytodienoic acid (OPDA) pools in Arabidopsis
Applicant Wolfender Jean-Luc
Number 163424
Funding scheme Project funding
Research institution Laboratoire de Pharmacognosie et Phytochimie Section des Sciences Pharmaceutiques Université de Genève
Institution of higher education University of Geneva - GE
Main discipline Biochemistry
Start/End 01.07.2016 - 31.12.2020
Approved amount 451'030.00
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All Disciplines (2)

Discipline
Biochemistry
Organic Chemistry

Keywords (13)

signalling; jasmonates; Arabidopsis thaliana; phytochemistry; wounding; UHPLC-TOF-MS; metabolomics; lipoxygenases LOX; oxylipins; defence; 12-oxo-phytodienoic acid; LC-MS/MS quantitation; plant defence

Lay Summary (French)

Lead
La réaction d’une plante à la blessure causée par un herbivore qui la ravage représente un processus clé de défense en écologie chimique. Les mécanismes qui régulent ce processus ont été beaucoup étudiés ces dernières années et il est notamment bien établi que la production de jasmonates (hormones de défense) est induite en réponse au stress causé par l’attaque des herbivores. Dans le cadre de ce projet nous essaierons de mieux comprendre les phénomènes qui régissent la mobilisation des précurseurs de ces jasmonates au moment de la blessure en utilisant Arabidopsis thaliana comme plante modèle.
Lay summary

Contenu et objectifs du travail de recherche

Lorsque des végétaux sont soumis à l’action d’herbivores, ils réagissent rapidement et fortement en induisant une réponse de défense largement régulée par les jasmonates. Ces hormones de défense sont dérivées d’acides gras insaturés qui sont convertis en précurseurs cyclopentenones (appelés ici (dn)OPDA).

La localisation cellulaire et la dynamique avec laquelle le (dn)OPDA) est mobilisé pour produire des jasmonates ou même son rôle doivent être élucidés. Afin d’étudier ceci en détail, nous allons utiliser une série de mutants d’Arabidopsis qui sont compromis dans leur défense et qui ne possèdent pas de lipoxygénase (13-LOX), une enzyme clé qui catalyse l’oxydation des acides gras. En étudiant ces mutants, il sera essentiel de comprendre quelle forme de (dn)OPDA) est exportée et recrutée dans la voie de biosynthèse de l’acide jasmonique. Pour ceci toute une série d’expériences sera menée avec les mutants 13-LOX et les niveaux de (dn)OPDA) seront mesurés très précisément par spectrométrie de masse. Nous pensons ainsi comprendre de façon détaillée le rôle de chacune de ces enzymes 13-LOX en lien avec la dynamique de production et la localisation cellulaire du (dn)OPDA). Ceci nous permettra d’obtenir un vue globale temporelle et spatiale de la dynamique de production et de recrutement du (dn)OPDA afin de comprendre au niveau moléculaire les premiers évènements qui déclenchent la réponse hormonale de défense à la blessure chez les végétaux.

 Contexte scientifique et sociétal du projet de recherche:

Ce projet devrait avoir un impact en agronomie notamment là où une compréhension détaillée des phénomènes de défense des plantes est importante dans le but de développer des stratégies de production de plantes de cultures.

Direct link to Lay Summary Last update: 21.05.2016

Responsible applicant and co-applicants

Employees

Publications

Publication
Wound‐ and mechanostimulated electrical signals control hormone responses
Farmer Edward E., Gao Yong‐Qiang, Lenzoni Gioia, Wolfender Jean‐Luc, Wu Qian (2020), Wound‐ and mechanostimulated electrical signals control hormone responses, in New Phytologist, 227(4), 1037-1050.

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
Developing Tools for Sustainable Crop Development Individual talk Newly identified components in long-distance wound signalling 27.05.2020 Durham, Great Britain and Northern Ireland Lenzoni Gioia;
Workshop Plants and Metabolomics Individual talk Strategies for unambiguous identification of biomarkers – Response to wounding in Arabidopsis thaliana as a case study 09.04.2019 Leiden, Netherlands Wolfender Jean-Luc;
Journée thématique SFSM – SFEAP : Spectrométrie de masse dans le monde du végétal Talk given at a conference Contextualized Metabolomics – A Paradigm Shift in Natural Product Research 04.04.2019 Paris, France Wolfender Jean-Luc;
International Regulatory Oxylipins meeting, VIB conferences Poster LOX6 isoform is necessary and sufficient for rapid long distance response after wounding in Arabidopsis thaliana. 01.04.2019 Ghent, Belgium Morin Hugo; Wolfender Jean-Luc; Lenzoni Gioia; Farmer Edward Elliston;
International Regulatory Oxylipins meeting, VIB conferences Talk given at a conference Mass spectrometry molecular networking a new tool for the profiling of specialized metabolites – Perspectives for studying the wound response in a holistic manner 01.04.2019 Ghent, Belgium Wolfender Jean-Luc; Morin Hugo; Farmer Edward Elliston;
International Regulatory Oxylipins meeting, VIB conferences Poster Identification of missing components in long-distance wound signalling 01.04.2019 Ghent, Belgium Lenzoni Gioia; Wolfender Jean-Luc; Farmer Edward Elliston; Morin Hugo;
Swiss Plant Molecular Biology Meeting Talk given at a conference Long-distance electrical signals in Arabidopsis leaves 30.03.2019 Meiringen, Switzerland Farmer Edward Elliston;
Organismic and Evolutionary Biology Talk given at a conference Long-Distance Electrical Signals in Arabidopsis Leaves 15.11.2018 Harvard , United States of America Farmer Edward Elliston;
Zermatt doctoral School of the School of Pharmaceutical Sciences Poster Mobilization and dynamics of 12-OXO phytodienoic acid (OPDA) resulting from early long distance wound signals in Arabidopsis thaliana. 01.08.2018 Zermatt, Switzerland Wolfender Jean-Luc; Morin Hugo;
Frontiers in Metabolomics Talk given at a conference Deep Metabolome Annotation - Challenges and Prospects in the Contextualization such Data for Bioactive Natural Product Prioritization 30.07.2018 Telluride, United States of America Wolfender Jean-Luc;
International Conference on Arabidopsis Research (ICAR) Talk given at a conference Vascular signalling routes for wound-induced jasmonate synthesis 25.06.2018 Turku, Finland Farmer Edward Elliston;
Metabolomics 2018 Talk given at a conference Contextualized Metabolomics Transforms Pharmacognosy – A Paradigm Shift in Natural Product Research 24.06.2018 Seattle, United States of America Wolfender Jean-Luc;
13èmes Rencontres Plantes-Bactéries Talk given at a conference How far deep can we get into the composition of natural extract metabolomes with modern mass spectrometry approaches? 29.01.2018 Aussois, France Wolfender Jean-Luc;
Keystone Conference ‘Plant Signalling’ Talk given at a conference Vascular Signaling Routes for Wound-Stimulated Jasmonate Synthesis 21.01.2018 Granlibakken, United States of America Farmer Edward Elliston;


Knowledge transfer events

Active participation

Title Type of contribution Date Place Persons involved
12th International Metabolomics Workshop Workshop 03.04.2020 Leiden, Netherlands Wolfender Jean-Luc;


Self-organised

Title Date Place

Associated projects

Number Title Start Funding scheme
146200 Targeted and non-targeted metabolomic study of the wound response in Arabidopsis: investigation of the role of 13-lipoxygenases in long distance signalling 01.04.2013 Project funding
183365 Ultra High Performance Liquid Chromatography-High Resolution Tandem Mass Spectrometry (UHPLC-HRMS/MS) for metabolomics and identification of bioactive molecules 01.10.2019 R'EQUIP
146200 Targeted and non-targeted metabolomic study of the wound response in Arabidopsis: investigation of the role of 13-lipoxygenases in long distance signalling 01.04.2013 Project funding
189921 An in silico and chemo-biological approach to identify anti-infective and pro-metabolic natural products 01.04.2020 Sinergia
164095 NMR 600MHz for metabolomics and biomarker identification for life sciences, chemical biology and medical projects at the University of Geneva 01.01.2016 R'EQUIP
205203 Mechanism of leaf-to-leaf electrical signalling in wounded plants 01.03.2022 Project funding

Abstract

When wounded by herbivores, plants mount a rapid and strong defence response that is regulated in large part by molecules known as jasmonates. These lipid regulators, the best known of which is the cyclopentanone jasmonic acid, derive from plastidic triunsaturated fatty acids that must first be converted to the 18-carbon cyclopentenone precursor oxo-phytodienic acid (OPDA) its 16-carbon homolog dinorOPDA (dnOPDA) (collectively named (dn)OPDA). Relatively little is known about the cellular localisations and dynamics of OPDA which possibly a signal in it's own right. However, some of this molecule is converted to jasmonates (JAs) upon wounding. Where does this take place and which pools of (dn)OPDA might be mobilised for JAs synthesis during attack? We have worked for several years on the dynamics of JAs pools in wounded plants. We now wish to tackle the technically more demanding aspect of studying (dn)OPDA dynamics. Our previous efforts have been focused on the development of metabolomics methods to discover novel forms of JAs and their dynamic in a robust wound model. More recently we have investigated the regulation of 13-lipoxygenases (13-LOXs), which are known to be involved in the upstream pathway of JA synthesis and we could highlight a paired hierarchical organization of the 13-LOXs in which LOX6-derived JAs help to coordinate the expression of other 13-LOXs. LOX2 was not found to be a major contributor to the expression of the key JAs-regulated rapid response gene JAZ10, but seems involved in producing high levels of arabidopsides. These latter compounds are esterified forms of OPDA and dnOPDA. In the present phase of the project we plan to specifically investigate the contribution of each 13-LOX to the pools of (dn)OPDA in free and bound forms. It is indeed essential to understand how and in which forms (dn)OPDA is exported from plastids and recruited in the pathway that leads to JAs production in Arabidopsis. We will especially perform experiments on triple mutants where all LOXs have been down-regulated with the exception of the LOX of interest. In such mutants we will precisely determine the levels of (dn)OPDA pools (free and bound) in resting plants and if lipid species containing (dn)OPDA are LOX-specific. We will use our wound model to investigate how fast bound pools of (dn)OPDA are mobilised and converted to JAs. The role of each specific (dn)OPDA pool will also be assessed in herbivory assays to understand if the bound forms may have a direct role in chemical defence. In addition we will conduct experiments to assess how (dn)OPDA is affected when expressing LOXs ectopically to shed light on the consequences of some 13-LOXs such as LOX6 having very specific cellular locations. For all these experiments we will use and further develop untargeted LC-HRMS general profiling methods for identifying all possible bound forms of (dn)OPDA and very sensitive HR-MS/MS for quantification of (dn)OPDA in all plant parts and specific tissues. We will also investigate state-of-the art FT-ICRMS imaging for localisation and sensitive NMR methods for general quantification of the galactolipid bound pools. Comprehensive temporal and spatial studies of (dn)OPDA pools dynamics in the 13-LOXs mutants using our wound model should give further insight of how (dn)OPDA is recruited in the pathway that lead to JAs and finally trigger wound signalling. Comparison of such events in non-producing arabidopside plants will also reveal what is unique about Arabidopsis and what can be generalised.
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