title: Analysis of fungal diversity of grapes by application of temporal temperature gradient gel electrophoresis - potentialities and limits of the method.
abstract: OBJECTIVE
  Fungi could be responsible for several problems in wines but the fungal ecosystem of grapes remains little known. The use of traditional methods does not allow to describe quickly this ecosystem. Therefore, we need to improve the knowledge about these fungi to prevent defects in wine. This study aims at evaluating the potentialities of the temporal temperature gradient gel electrophoresis (TTGE) method to describe the fungal ecosystem of grapes.
  
  
  RESULTS
  The internal transcribed spacer (ITS) region was amplified and analysed using TTGE. A reference database of 56 fungal species was set up to evaluate the discrimination power of the method. The database was used for the direct identification of the fungal species present in complex samples. The sensitivity of the method is below 10(4) spores per ml.
  
  
  CONCLUSIONS
  This method allows to describe the fungal diversity of grapes, but does not always allow to directly identify all fungal species, because of the taxonomic resolution of the ITS sequences. However, this identification strategy is less time consuming than traditional analysis by cloning and sequencing the bands.
  
  
  CONCLUSIONS
  With this method, it will be possible to compare the fungal species present in different vineyards and to connect the presence of some fungi with particular defects in wine.
authors.affiliation: Département Microbiologie Industrielle, Ecole Nationale Supérieure des Industries Agricoles et Alimentaires, Massy Cedex, France.
authors.forename: E
authors.initials: E
authors.lastname: Doaré-Lebrun
authors.forename: A
authors.initials: A
authors.lastname: El Arbi
authors.forename: M
authors.initials: M
authors.lastname: Charlet
authors.forename: L
authors.initials: L
authors.lastname: Guérin
authors.forename: J-J
authors.initials: JJ
authors.lastname: Pernelle
authors.forename: J-C
authors.initials: JC
authors.lastname: Ogier
authors.forename: M
authors.initials: M
authors.lastname: Bouix
mesh_terms: D004271:DNA, Fungal / Q000032:analysis; D030541:Databases, Genetic; D004591:Electrophoresis, Polyacrylamide Gel; D005658:Fungi / Q000235:genetics*; D014644:Genetic Variation; D007218:Industrial Microbiology; D013170:Spores; D027843:Vitis / Q000382:microbiology*
chemical_list: D004271:DNA, Fungal