Detection of Dekkera-Brettanomyces strains in sherry by a nested PCR method.
Brettanomyces sp. and its ascosporogenous sexual state, Dekkera sp., have been well documented as spoilage microorganisms, usually associated with barrel-aged red wines. In this report, we describe the genetic characterization, on the basis of DNA content per cell, electrophoretic karyotyping, and mitochondrial DNA restriction patterns, of a Dekkera yeast strain isolated from sherries and of a number of other Brettanomyces and Dekkera strains. By using a genomic DNA fragment of the isolated Dekkera strain, we developed a two-step PCR method which directs the specific amplification of target DNA from this strain and from other Brettanomyces-Dekkera strains. The method efficiently amplified the target DNA from intact cells, obviating DNA isolation, and yielded a detection limit of fewer than 10 yeast cells in contaminated samples of sherry.
Ibeas
J I
JI
Unidad de Genetica, Facultad de Ciencias, Universidad de Malaga, Campus Universitario de Teatinos, Spain.
Lozano
I
I
Perdigones
F
F
Jimenez
J
J
D001483:Base Sequence; D004271:DNA, Fungal / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D014920:Wine / Q000382:microbiology*; D015003:Yeasts / Q000235:genetics / Q000302:isolation & purification*
D004271:DNA, Fungal