Development of a molecular method for the typing of Brettanomyces bruxellensis (Dekkera bruxellensis) at the strain level.
OBJECTIVE
In recent years, Brettanomyces/Dekkera bruxellensis has caused increasingly severe quality problems in the wine industry. A typing method at the strain level is needed for a better knowledge of the dispersion and the dynamics of these yeasts from grape to wine.


RESULTS
Three molecular tools, namely random-amplified polymorphic DNA, PCR fingerprinting with microsatellite oligonucleotide primers and SAU-PCR, were explored for their relevance to typing strains of Brettanomyces bruxellensis. The results indicated that discrimination of each individual strain was not possible with a single PCR typing technique. We described a typing method for B. bruxellensis based on restriction enzyme analysis and pulse field gel electrophoresis (REA-PFGE). Results showed that electrophoretic profiles were reproducible and specific for each strain under study.


CONCLUSIONS
Consequently, REA-PFGE should be considered for the discrimination of B. bruxellensis strains. This technique allowed a fine discrimination of B. bruxellensis, as strains were identified by a particular profile.


CONCLUSIONS
This study constitutes a prerequisite for accurate and appropriate investigations on the diversity of strains throughout the winemaking and ageing process. Such studies will probably give clearer and more up-to-date information on the origin of the presence of Brettanomyces in wine after vinification when they are latent spoilage agents.
Miot-Sertier
C
C
UMR Oenologie-Ampélologie, INRA-Faculté d'Oenologie, Université Victor Segalen Bordeaux 2, Talence Cedex, France.
Lonvaud-Funel
A
A
D016172:DNA Fingerprinting / Q000379:methods; D004271:DNA, Fungal / Q000032:analysis*; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods; D005506:Food Contamination; D005516:Food Microbiology; D018895:Microsatellite Repeats; D016533:Mycological Typing Techniques; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods; D013045:Species Specificity; D014920:Wine; D015003:Yeasts / Q000235:genetics*
D004271:DNA, Fungal