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J Microbiol Methods


Title:Identification and profiling of volatile metabolites of the biocontrol fungus Trichoderma atroviride by HS-SPME-GC-MS
Author(s):Stoppacher N; Kluger B; Zeilinger S; Krska R; Schuhmacher R;
Address:"Center for Analytical Chemistry, Department for Agrobiotechnology (IFA-Tulln), University of Natural Resources and Applied Life Sciences, Vienna, Konrad Lorenz Strasse 20, A-3430 Tulln, Austria"
Journal Title:J Microbiol Methods
Year:2010
Volume:20100316
Issue:2
Page Number:187 - 193
DOI: 10.1016/j.mimet.2010.03.011
ISSN/ISBN:1872-8359 (Electronic) 0167-7012 (Linking)
Abstract:"In the present study we describe a method, which is based on solid phase microextraction (SPME) coupled to gas chromatography-mass spectrometry (GC-MS) and which can be used for the profiling of microbial volatile organic compounds (MVOCs) in the headspace (HS) of cultures of filamentous fungi. The method comprises the following successive steps: 1. growth of the fungus on a solid culture medium directly in headspace vials, 2. measurement of volatiles by HS-SPME-GC-MS, 3. deconvolution of mass spectra, 4. identification of volatiles by comparison of measured, deconvoluted mass spectra and linear temperature programmed retention indices (LTPRI) on two stationary GC phases with database entries and LTPRI published in the literature, and 5. profiling of the identified MVOCs. The developed method was successfully applied to cultures of the biocontrol fungus Trichoderma atroviride. An in-house library consisting of mass spectra and LTPRI values of fungal VOCs was established and used to study the profiles of MVOCs of this fungus. In total, 25 different MVOCs were identified by applying strict criteria (spectral match factor at least 90% and a maximum relative deviation of LTPRI of +/-2% from literature values). The MVOCs were assigned to the compound classes of alcohols, ketones, alkanes, furanes, pyrones (mainly the bioactive 6-pentyl-alpha-pyrone), mono- and sesquiterpenes, 13 of which have never been reported to be produced by Trichoderma spp. before. Eleven of these volatiles have been additionally confirmed using authentic standards. Finally, time course experiments and cultivation of T. atroviride in the presence of the mycotoxin fusaric acid demonstrated the potential of the method to study the dynamics of MVOC profiles as well as the effect of different environmental/biological conditions on the expression of MVOCs of filamentous fungi"
Keywords:Gas Chromatography-Mass Spectrometry *Metabolome Solid Phase Microextraction Trichoderma/*chemistry/growth & development/metabolism Volatile Organic Compounds/*analysis;
Notes:"MedlineStoppacher, Norbert Kluger, Bernhard Zeilinger, Susanne Krska, Rudolf Schuhmacher, Rainer eng Evaluation Study Research Support, Non-U.S. Gov't Netherlands 2010/03/23 J Microbiol Methods. 2010 May; 81(2):187-93. doi: 10.1016/j.mimet.2010.03.011. Epub 2010 Mar 16"

 
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