6533b851fe1ef96bd12a8f31
RESEARCH PRODUCT
Species-specific identification of Dekkera/Brettanomyces yeasts by fluorescently labeled DNA probes targeting the 26S rRNA.
Christoph RöderJürgen FröhlichHelmut Königsubject
Sequence analysisBrettanomycesMolecular Sequence DataWineBiologyApplied Microbiology and BiotechnologyMicrobiologySpecies SpecificityDNA FungalMycological Typing TechniquesIn Situ Hybridization FluorescencePhylogenyDNA PrimersGeneticsBase SequenceHybridization probeFungal geneticsNucleic acid sequenceGeneral MedicineSequence Analysis DNARibosomal RNAbiology.organism_classificationYeastNucleic Acid ProbesRNA RibosomalSaccharomycetalesNucleic Acid ConformationSpecific identificationdescription
Sequencing of the complete 26S rRNA genes of all Dekkera/Brettanomyces species colonizing different beverages revealed the potential for a specific primer and probe design to support diagnostic PCR approaches and FISH. By analysis of the complete 26S rRNA genes of all five currently known Dekkera/Brettanomyces species (Dekkera bruxellensis, D. anomala, Brettanomyces custersianus, B. nanus and B. naardenensis), several regions with high nucleotide sequence variability yet distinct from the D1/D2 domains were identified. FISH species-specific probes targeting the 26S rRNA gene's most variable regions were designed. Accessibility of probe targets for hybridization was facilitated by the construction of partially complementary 'side'-labeled probes, based on secondary structure models of the rRNA sequences. The specificity and routine applicability of the FISH-based method for yeast identification were tested by analyzing different wine isolates. Investigation of the prevalence of Dekkera/Brettanomyces yeasts in the German viticultural regions Wonnegau, Nierstein and Bingen (Rhinehesse, Rhineland-Palatinate) resulted in the isolation of 37 D. bruxellensis strains from 291 wine samples.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2007-06-29 | FEMS yeast research |