Evaluation of fungal and yeast diversity in Slovakian wine-related microbial communities |
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Authors: | Barbara Bre?ná Katarína ?eni?ová Katarína Chovanová Viera Chebeňová Lucia Kraková Tomá? Kuchta Domenico Pangallo |
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Institution: | 1.Food Research Institute,Bratislava,Slovak Republic;2.Institute of Molecular Biology,Slovak Academy of Sciences,Bratislava,Slovak Republic;3.Caravella s.r.o,Bratislava,Slovak Republic |
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Abstract: | Since the yeast flora of Slovakian enology has not previously been investigated by culture-independent methods, this approach
was applied to two most common cultivars Frankovka (red wine) and Veltlin (white wine), and complemented by cultivation. Model
samples included grapes, initial must, middle fermenting must and must in the end-fermentation phase. The cultured isolates
were characterized by length polymorphism of rDNA spacer two region using fluorescence PCR and capillary electrophoresis (f-ITS
PCR), and some were identified by sequencing. The microbial DNA extracted directly from the samples without cultivation was
analysed by f-ITS PCR, amplicons were cloned and sequenced. The use of universal fungal primers led to detection of both yeasts
and filamentous fungi. The amplicon of highest intensity and present in all the samples corresponded to Hanseniaspora uvarum. Other species demonstrated by both approaches included Saccharomyces sp., Metschnikowia pulcherrima or M. chrysoperlae, Candida zemplinina, Cladosporium cladosporioides, Botryotinia fuckeliana, Pichia anomala, Candida railenensis, Cryptococcus magnus, Metschnikowia viticola or Candida kofuensis, Pichia kluyveri or Pichia fermentas, Pichia membranifaciens, Aureobasidium pullulans, Alternaria alternata, Erysiphe necator, Rhodotorula glutinis, Issatchenkia terricola and Debaryomyces hansenii. Endemism of Slovakian enological yeasts was suggested on the level of minor genetic variations of the known species and probably
not accounting for novel species. The prevalence of H. uvarum over Saccharomyces sp. in the samples was indicated. This is the first culture-independent study of Slovakian enology and the first time f-ITS
PCR profiling was used on wine-related microbial communities. |
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