New PCR-based methods for yeast identification.
OBJECTIVE
To characterize reference yeast strains and identify indigenous strains isolated from wine fermentations by PCR methods.


RESULTS
We compared several PCR techniques for yeast identification. We used oligonucleotide primers that are complementary to (i) intron splice sites, (ii) REP and (iii) ERIC elements to produce PCR fingerprints that display specific patterns between the different yeast species. These three techniques were used to characterize 41 reference yeast strains belonging to 15 different species and to identify 40 indigenous strains isolated from grape must and wine fermentations. Species-specific banding patterns were obtained with the three PCR-techniques with different degrees of intraspecific differentiation depending on the method. By comparing the PCR fingerprints of unknown isolates with those produced by reference strains, we identified yeast strains isolated from an industrial wine fermentation.


CONCLUSIONS
All three PCR techniques are rapid, reliable and simple methods of yeast identification. As far as we know, this is the first time that the primers designed for amplifying repetitive elements in bacteria have been successfully used in yeast.


CONCLUSIONS
Industry needs rapid, reliable and simple methods of yeast identification. The proposed PCR techniques will allow to achieve this objective.
Hierro
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N
Departament de Bioquímica i Biotecnologia, Facultat d'Enologia, Unitat d'Enologia, Centre de Referència en Tecnologia d'Aliments (CeRTA), Universitat Rovira i Virgili, Ramón y Cajal, Tarragona, Spain.
González
A
A
Mas
A
A
Guillamón
J M
JM
D016172:DNA Fingerprinting / Q000379:methods; D004271:DNA, Fungal / Q000032:analysis; D005285:Fermentation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA / Q000379:methods; D013045:Species Specificity; D014920:Wine / Q000382:microbiology*; D015003:Yeasts / Q000302:isolation & purification*
D004271:DNA, Fungal