Typing of Saccharomyces cerevisiae and Kloeckera apiculata strains from Aglianico wine

Abstract

Aims: Kloeckera apiculata and Saccharomyces cerevisiae yeast species are dominant, respectively, at the early and at the following stages of wine fermentation. In the present study, PCR fingerprinting and NTS region amplification and restriction were applied as techniques for monitoring yeast population performing Aglianico of Vulture grape must fermentation. Methods and Results: Thirty S. cerevisiae and 30 K. apiculata strains were typed by PCR fingerprinting with (GAC)5 and (GTG)5 primers and by complete NTS region amplification followed by restriction with HaeIII and MspI enzymes. S. cerevisiae strains generated two patterns with (GAC)5 primer, while (GTG)5 primer yielded a higher genetic polymorphism. Conversely, in K. apiculata Aglianico wine strains (GAC)5 and (GTG)5 primers generated the same profile for all strains. Restriction analysis of the amplified NTS region gave the same profile for all strains within the same species, except for one strain of S. cerevisiae. Conclusions: The PCR fingerprinting technique was useful in discriminating at strain level S. cerevisiae, particularly with the primer (GTG)5. RFLP patterns generated from the NTS region of the two species can be more easily compared than the patterns resulting from PCR fingerprinting, thus RFLP is more suitable for the rapid monitoring of the species involved in different stages of fermentation. Significance and Impact of the Study: The molecular techniques used allow discrimination of S. cerevisiae at strain level and monitoring of the ratio of S. cerevisiae/K. apiculata during the fermentation process. Thus, their application can assure technological adjustments in a suitable time.

Biogeography, Host Specificity, and Molecular Phylogeny of the Basidiomycetous Yeast Phaffia rhodozyma and Its Sexual Form, Xanthophyllomyces dendrorhous

“…MSP-PCR fingerprinting has been used to type fungal strains by several research groups. In most cases species-specific PCR patterns are obtained (17,25,29), but in some instances this approach results in intraspecific differentiation (4,18). Our results suggest that primers (GTG) 5 and M13 are useful to discriminate populations of Phaffia/ Xanthophyllomyces.…”

Cryptococcus terrestris sp. nov., a tremellaceous, anamorphic yeast phylogenetically related to Cryptococcus flavescens

“…3). Fingerprinting with this primer usually gives species-specific patterns, but there are other examples of pattern differentiation among geographically separated strains belonging to the same species (Caruso et al, 2002;Libkind et al, 2007). In spite of the different fingerprinting profiles, the identical phenotypic characteristics and the D1/D2 and ITS sequences indicate that the Oklahoman and Brazilian isolates belong to the same species.…”

Molecular Identification and Characterization of Wine Yeasts

“…This is the case for the NTS region (Baleiras Couto et al, 1996;Capece et al, 2003;Caruso et al, 2002;Nguyen & Gaillardin, 1997;Pulvirenti et al, 2000), the 18S gene (Capece et al, 2003), and various domains of the 26S gene (Baleiras Couto et al, 1996Romancino et al, 2008;SmoleMozina et al, 1997;van Keulen et al, 2003). However, the absence of a sequence database for these regions means that their use cannot be generalized for the identification of yeasts.…”

“…Studies have also been undertaken to analyze molecular variation in natural populations of S. cerevisiae using restriction analysis of mtDNA (Granchi et al, 2003;Gutiérrez et al, 1997;Pramateftaki et al, 2000;Querol et al, 1994;Sabaté et al, 1998;Torija et al, 2001), electrophoretic karyotyping (Frezier & Dubourdieu, 1992;Schü tz & Gafner, 1993, microsatellite analysis (Caruso et al, 2002;Howell et al, 2004), fluorescent in situ hybridization (FISH) (Xufre et al, 2006), or a combination of techniques (Egli et al, 1998;Ló pes et al, 2002;Nadal et al, 1996;Rodríguez et al, 2004;Torriani et al, 1999;Versavaud et al, 1995). These studies have shown that different strains of S. cerevisiae dominate fermentation in different appellations.…”