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Plant metabolite analysis by liquid chromatography-mass spectrometry

English title Plant metabolite analysis by liquid chromatography-mass spectrometry
Applicant Martinoia Enrico
Number 145020
Funding scheme R'EQUIP
Research institution Institut für Pflanzen- und Mikrobiologie Universität Zürich
Institution of higher education University of Zurich - ZH
Main discipline Botany
Start/End 01.10.2013 - 30.09.2014
Approved amount 250'000.00
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All Disciplines (4)

Discipline
Botany
Ecology
Cellular Biology, Cytology
Biochemistry

Keywords (6)

herbivore; High Pressure Liquid Chromatography-Mass Spectrome; Phenolics; Secondary metabolites; Chlorophyll; Phytohormones

Lay Summary (German)

Lead
Beschaffung eines neuen HPLC-MS (High Pressure Liquid Chromatography-Mass Spectrometry) Geräts am Standort Botanischer Garten unterstützt
Lay summary

Lebende Organismen führen eine Vielzahl biochemischer Reaktionen aus, die unter zellulärem Metabolismus zusammen gefasst werden können. Diese Reaktionen erlauben den Organismen zu wachsen und ihre Form auszubilden, sich zu vermehren und adäquat auf die verschiedenen Umwelteinflüsse zu reagieren. Daher ist die Produktion und der Abbau von Metaboliten ein sehr dynamischer und komplexer Prozess. In den letzten Jahren wurden hoch sensible Techniken entwickelt, um auch Metabolite die in sehr kleinen Mengen vorkommen, zu messen. Diese stark verbesserten Analysemethoden erlauben einerseits fokussiert eine definierte Substanz zu bestimmen, anderseits erlauben sie einen so genannten “non-targeted” Ansatz durchzuführen, indem in einer Probe möglichst die Gesamtheit der Substanzen aufgetrennt und analysiert wird. Die Gesamtheit der bestimmten Substanzen wird Metabolom genannt. Das durch den Schweizerischen Nationalfonds und die Universität Zürich gemeinsam finanzierte Flüssigchromatographie-Massenspektrometrie Gerät wird in unserem Institut verwendet werden, um für den Projekte, die an unserem Institut bearbeitet werden, neue Einsichten in metabolische Prozesse zu erhalten. Dazu gehören die Aufklärung des Chlorophyllabbaus, der Transport von Ligninvorläufermolekülen, die Untersuchung der Veränderungen der Metabolite bei Befall verschiedener Pathogene von Weizen und Gerste, die Bestimmung von Phytohormonen und pflanzlichen Sekundärmetaboliten, sowohl unter verschiedenen Umweltbedingungen als auch in der Insekten-Pflanze Interaktion. Wir werden dieses Gerät daher für die gezielte Analyse von Substanzen als auch für den „non-targeted“ Ansatz verwenden und erhoffen uns dadurch grosse, neue Erkenntnisse für unsere Projekte

Direct link to Lay Summary Last update: 07.03.2013

Responsible applicant and co-applicants

Collaboration

Group / person Country
Types of collaboration
Harro Bouwmeester, University Wageningen Netherlands (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Alisdair Fernie, Max-Planck Institute for Molecular Plant Physiology, Golm Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Laurent Bigler, Institute of Organic Chemistry, University of Zurich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Exchange of personnel
Bernhard Kräutler; University of Innsbruck Austria (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Exchange of personnel

Associated projects

Number Title Start Funding scheme
132603 Chlorophyll breakdown: analysis of fruit ripening, evolution and regulation 01.01.2011 Project funding (Div. I-III)
135747 ABC transporters involved in signalling events 01.04.2011 Project funding (Div. I-III)
138472 The TOR (Target of Rapamycin) signalling pathway - a new modulator of cell wall development 01.02.2012 Project funding (Div. I-III)
134936 Structure, function and biosynthesis of syrbactins 01.01.2012 Project funding (Div. I-III)
127021 Spatio-Temporal Memory Streaming 01.09.2010 Project funding (Div. I-III)
134416 InvaVol: What are the consequences of herbivore attack for floral signals? 01.04.2011 Project funding (special)

Abstract

Living organisms, including plants, perform a multitude of biochemical reactions, in summary termed their metabolism. These reactions enable an organism to grow and maintain its structure, to reproduce and respond to its environment. Therefore, metabolic processes need to be coordinated with respect to both time and space, and occurrence of metabolites, i.e. the intermediates and products of metabolism, is highly dynamic and complex. The analysis of metabolites within a given organism/organ/cell faces two major challenges: (i) metabolite concentrations can vary by orders of magnitude, and (ii) metabolites can extremely differ in their chemical properties. A widely-used separation technique for metabolite analysis is liquid chromatography (LC), in particular high performance liquid chromatography (HPLC). To reach high sensitivity of compound detection and for mass and/or structure determination of a metabolite sensitive analytical methods like mass spectrometry (MS) are required. Technical developments within the last years considerably improved sensitivity, flexibility and user-friendliness of respective equipment, allowing a wide range of LC-MS technologies to become state-of-the-art in non-specialized environments or laboratories.Accordingly, metabolomics is a rapidly developing field in the post-genomic era and major progress has been made in both non-targeted and targeted metabolite analysis in different research areas, including plant science. LC-MS-based targeted metabolomic approaches have been widely used in plant research to analyze both primary and secondary metabolites, ranging from hydrophilic to very hydrophobic compound classes. Examples are plant lipids, isoprenoids, hormones, flavonols, pigments, and amino acids. The equipment for metabolite analysis currently available at the Institute of Plant Biology at the University of Zurich is outdated and does no longer meet our requirements for high-resolution and high-throughput metabolite analyses. Because of this deficiency, complex, high throughput and particularly sensitive metabolite analyses in our research largely depend on external collaborations. Although these collaborations are extremely successful, as seen from many joint publications, our collaborating groups often do not have the capacity and sufficient time to fully meet our demands. Furthermore, the geographical distance of several of our collaborators within and outside Switzerland make the use of the external equipment inapplicable for its use on a (nearly) daily basis as required in our projects. In summary, these constraints considerably block the progress of our research. To maintain our high scientific competence and competitiveness in the areas of our research where metabolite analysis is fundamental, we would need our own state-of-the-art LC-MS equipment. The system we are applying for in this R’Equip proposal would meet our requirements for high-resolution and high-throughput metabolite analyses needed for many projects currently studied at the Institutes of Plant Biology and Systematic Botany. These include:•Transport of monolignols•Establishing protocols for measuring phytohormones and their conjugates•Non-targeted approaches to identify new vacuolar transporters and transporters involved in secretion of secondary compounds•Characterization of grape berries anthocyanin transporters•Identification and analysis of metabolites of chlorophyll breakdown•Metabolic profiling of leaf rust resistance in cereal crops•Structural analysis of syrbactin-type compounds•Flavonol accumulation profiling in Arabidopsis thaliana•Pollinator- and herbivore-mediated selection on plant secondary metabolites
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