EFS12- Book of abstracts - Contact
EFS12- Book of abstracts - Contact
EFS12- Book of abstracts - Contact
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SESSION 2: SECONDARY METABOLITES – BIOCHEMISTRY,<br />
BIOSYNTHESIS, FEED AND FOOD SAFETY<br />
Fusarium graminearum in depth: a novel method to<br />
identify new metabolites by isotopic labelling and<br />
high resolution mass spectrometry<br />
P. M. Cano 1,2 , E. Jamin 1 , S. Tadrist 1 , P. Bourdaudhui 1 , M. Péan 3,4,5 , L.<br />
Debrauwer 1 , I. P. Oswald 1,2 , M. Delaforge 6 , O. Puel 1,2<br />
1 INRA, UMR 1331, Toxalim, Research Center in Food Toxicology, F-31027 Toulouse, France;<br />
2 Université de Toulouse, INP, Toxalim, F-31076 Toulouse, France; 3 CEA, DSV, IBEB, Groupe de<br />
Recherches Appliquées en Phytotechnologie, F-13108 Saint-Paul-les-Durance, France; 4 CNRS, UMR<br />
Biologie Végétale & Microbiologie Environnementale, F-13108 Saint-Paul-les-Durance, France; 5 Aix-<br />
Marseille Université, F-13108 Saint-Paul-les-Durance, France; 6 CEA Saclay, iBiTec-S, SB2SM and<br />
URA CNRS 8221, F-91191 Gif sur Yvette, France<br />
E-mail: paty.canog@gmail.com<br />
Characterization <strong>of</strong> fungal secondary metabolomes has become a challenge <strong>of</strong><br />
great interest in the last decades due to the emergence <strong>of</strong> fungal threats to<br />
natural ecosystems and public health; and also due to the industrial interest <strong>of</strong><br />
many <strong>of</strong> these molecules. In view <strong>of</strong> this, the aim <strong>of</strong> the present study was to<br />
develop an integrated approach to analyse fungal metabolomes. The method we<br />
present hereby combines high resolution mass spectrometry and double isotopic<br />
labelling which efficiently enabled the unambiguous determination <strong>of</strong> exact<br />
chemical formulas, getting rid <strong>of</strong> problems coming from interference <strong>of</strong> nonbiological<br />
molecules. More precisely, the Aspergillus fumigatus strain NRRL<br />
35693, an extremely hazardous human pathogen and the Fusarium graminearum<br />
strain PH1, a devastating plant pathogen, were grown on wheat grains (Triticum<br />
aestivum) with different isotopic enrichments: (1) naturally enriched grains, (2)<br />
grains enriched 96.88% 13 C, (3) grains enriched with 53.37% 13 C and 96.8% 15 N.<br />
Methanol extracts <strong>of</strong> each culture was then analysed by reversed phase liquid<br />
chromatography coupled to LTQ-Orbitrap mass spectrometer. Data <strong>of</strong> the 3<br />
cultures were cross-analysed with an in-house developed s<strong>of</strong>tware. Metabolites<br />
were characterized with the metabolite database, Antibase 2012, annotated with<br />
MS/MS experiments and identified by comparison with standards when possible.<br />
The method was firstly successfully validated with the well-known metabolome <strong>of</strong><br />
Aspergillus fumigatus. Application <strong>of</strong> the method on the metabolome <strong>of</strong> Fusarium<br />
graminearum allowed the characterization <strong>of</strong> 37 new compounds including<br />
fusaristatin A which had never been isolated from this specie before, bringing a<br />
new perspective on the toxicity <strong>of</strong> this fungus. This kind <strong>of</strong> analysis will<br />
undoubtedly facilitate the study <strong>of</strong> fungal metabolomes.<br />
Keywords: Fusarium graminearum, metabolome, isotopic labelling, HPLC-FT-MS<br />
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