<div class="csl-bib-body">
<div class="csl-entry">Kirnbauer, S., Missbach, K., Flatschacher, D., Schuhmacher Rainer, Zeilinger Susanne, & Marchetti-Deschmann, M. (2022, September 2). <i>Monitoring Metabolite Distributions During the interaction of T.atroviride and B.cinerea by MALDI FTICR MS Imaging,</i> [Conference Presentation]. IMSC 2022, Maastricht, Netherlands (the). http://hdl.handle.net/20.500.12708/152722</div>
</div>
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dc.identifier.uri
http://hdl.handle.net/20.500.12708/152722
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dc.description.abstract
study aims for a better understanding of underlying mechanisms allowing T.atroviride
to actually sense a pathogen. For this we developed an agar-based micro assay to study
especially the formation of secondary metabolites (SMs) that play a crucial role in fungal
sensing and interaction. Applying MALDI MSI on an 7T FT-ICR instrument we monitor the
spatial distribution of known and unknown substances in an interaction zone (almost) not
harboring fungal hyphae. By this we aim to identify highly mobile SMs migrating towards
the interaction partner resulting in a response in the interaction partners T.atroviride and
B.cinerea.
Methods
Pre-cultures grown at 25°C in petri dishes were transferred on opposite ends of an agar medium
deposited on glass or ITO slides. Incubation was performed until growth fronts
were 1cm apart (confrontation zone). After quenching (liquid nitrogen) and vacuum drying,
matrix deposition was done with a home-built sublimation unit or a TM sprayer
(HTX). Extractions of the confrontation zone were done with acidified water/acetonitrile.
MALDI MSI of the confrontation zone and direct-infusion ESI was performed on a 7T
scimaX FT-ICR-MRMS equipped with an ESI/MALDI dual source (Bruker). Data analysis
was done with SCiLS and DataAnalysis 5.0 (Bruker).
Preliminary data (results)
A confrontation assay of standardized measures was developed for regular glass or ITO
slides. This standardization in preparation makes parameters like fungal growth highly
reproducible. The miniaturization on commercially available glass slides also allows for
samples measurement by MALDI directly after quenching, drying and matrix application
using commercial sample holders.
The distribution of peptaibols and other metabolites formed by T. atroviride as well as
potentially novel metabolites from B.cinerea and T. atroviride were visualized by MALDI
MSI in the confrontation zone.
The agar is producing high chemical backgrounds for every MSI experiment. To
overcome this limitation of our approach we tested different matrices (e.g. 2,5-DHB,
CHCA, 1,5-DAN, 3-Hydroxycoumarin, 6-Aza-2-Thiothymine and mixtures thereof) for
efficiency and most comprehensive analysis of the analyte signals we can detect. Furthermore, sublimation
and spraying for matrix deposition were studied in more detail for agar samples and
experiments for derivatization of functional groups were conducted to improve the signal-to-noise ratio of interesting target compounds.
To confirm our findings, the agar from the confrontation zone was extracted with a
mixture of 50:50 (v:v) ACN:H2O acidified with 0.1% Formic acid or 0.1% Acetic acid.
Direct-infusion ESI FT-ICR MS analyses allowed further evaluation of ESI versus MALDI
ionization efficiencies of metabolites to assess the potential for detecting metabolites in
the presented MALDI MSI setup. Our putative results were then compared with
metabolomics outcomes from experiments using isotopically-labeled standards in the
same assay to verify the fungal origin of the metabolites.
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dc.description.sponsorship
Fonds zur Förderung der wissenschaftlichen Forschung (FWF)
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dc.language.iso
en
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dc.subject
Mass Spectrometry, Imaging, Microbiology
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dc.title
Monitoring Metabolite Distributions During the interaction of T.atroviride and B.cinerea by MALDI FTICR MS Imaging,
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dc.type
Presentation
en
dc.type
Vortrag
de
dc.contributor.affiliation
Universität Innsbruck, Austria
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dc.relation.grantno
P32179-B
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dc.type.category
Conference Presentation
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tuw.project.title
Chemische Kommunikation in mykoparasitischen Interaktionen
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tuw.researchTopic.id
M6
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tuw.researchTopic.name
Biological and Bioactive Materials
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tuw.researchTopic.value
100
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tuw.publication.orgunit
E164-01-1 - Forschungsgruppe Massenspektrometrische Bio- und Polymeranalytik
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tuw.author.orcid
0000-0003-2202-1593
-
tuw.author.orcid
0000-0001-8221-9695
-
tuw.author.orcid
0000-0002-7520-4943
-
tuw.author.orcid
0000-0003-3112-0948
-
tuw.author.orcid
0000-0002-8060-7851
-
tuw.event.name
IMSC 2022
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tuw.event.startdate
27-08-2022
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tuw.event.enddate
02-10-2022
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tuw.event.online
On Site
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tuw.event.type
Event for scientific audience
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tuw.event.place
Maastricht
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tuw.event.country
NL
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tuw.event.institution
IMSF
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tuw.event.presenter
Kirnbauer, Stefan
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tuw.event.track
Multi Track
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wb.sciencebranch
Chemie
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wb.sciencebranch.oefos
1040
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wb.sciencebranch.value
100
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item.fulltext
no Fulltext
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item.openairetype
conference paper not in proceedings
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item.languageiso639-1
en
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item.grantfulltext
none
-
item.openairecristype
http://purl.org/coar/resource_type/c_18cp
-
item.cerifentitytype
Publications
-
crisitem.author.dept
E164-01-1 - Forschungsgruppe Massenspektrometrische Bio- und Polymeranalytik
-
crisitem.author.dept
BOKU University
-
crisitem.author.dept
Universit�t Innsbruck
-
crisitem.author.dept
E164 - Institut für Chemische Technologien und Analytik
-
crisitem.author.orcid
0000-0003-2202-1593
-
crisitem.author.orcid
0000-0001-8221-9695
-
crisitem.author.orcid
0000-0002-8060-7851
-
crisitem.author.parentorg
E164-01 - Forschungsbereich Imaging und Instrumentelle Analytische Chemie
