Use of interdelta polymorphisms of Saccharomyces cerevisiae strains to monitor population evolution during wine fermentation

Abstract

The industrial use of starter cultures containing a consortium of different strains from the same species is nowadays seen as a possible strategy to enhance the organoleptic complexity of wines. To assess the relative contribution of each strain to the final product it is essential to quantify population evolution during the wine fermentation process, which requires strain-specific methods to identify and differentiate each strain. In the present study, a molecular method based on analysis of the polymorphisms exhibited by the PCR-amplification of the delta regions of three Saccharomyces cerevisiae strains was developed. A set of three pairs of primers (delta1-delta2, delta12-delta2, delta12-delta21) was used for each strain, and analysis of the resulting polymorphism patterns showed that the delta12-delta2 primer pair exhibited the highest resolution and discriminatory power. Thus, this pair of primers was selected to monitor the population evolution of a laboratory-scale wine fermentation performed in synthetic grape juice that was inoculated with similar amounts of each strain. The results showed that all strains grew together during the exponential growth phase (2-3 days) and maintained high cell density values (10(6)-10(7) cfu ml(-1)) throughout the stationary growth phase without significantly changing their relative population proportion, thus indicating that each strain can influence the chemical composition and final flavor of wine, albeit at different levels. This study also showed that PCR-amplification of DNA delta sequences of S. cerevisiae strains is a reproducible, strain-specific and simple method that can be used successfully to monitor yeast strain population dynamics during wine fermentations.

Autochthonous fermentation starters for the industrial production of Negroamaro wines

“…The selected starter cultures could hereafter be applied to better investigate the use of industrial starters composed of a consortium of either S. cerevisiae [63] or Saccharomyces and non-Saccharomyces mixed strains [7,8]. These approaches are nowadays considered as possible strategies to enhance the organoleptic complexity of wines, which is naturally produced by the complex microbial process operated by cultivable and non-cultivable microbial species during the winemaking process [41].…”

Survival rate of wine-related yeasts during alcoholic fermentation assessed by direct live/dead staining combined with fluorescence in situ hybridization

“…During the last decades, molecular techniques such as AFLP, microsatellites, RAPD, DGGE and interdelta polymorphisms have been used, either directly or after plate isolation, to identify and differentiate wine yeast species and strains (De Barros Lopes et al, 1999;Maqueda et al, 2010;Prakitchaiwattana et al, 2004;Xufre et al, 2000Xufre et al, , 2011. More recently, direct molecular methods such as quantitative PCR (qPCR) have been used to quantify microbial species and to monitor wine fermentations (Andorrà et al, 2008;Hierro et al, 2006Hierro et al, , 2007.…”

Morphophysiological and molecular characterization of wild yeast isolates from industrial ethanol process

“…This can be observed in the results obtained by the analysis of 26S rDNA sequencing (Figure 1), where isolates presented distinct banding patterns when compared to the reference strains. This can be caused by the emergence of a native strain in the process, or due to adaptive evolution, as mentioned by Argueso et al (2009), Burke (2012) and Xufre et al (2011). The results made it possible to identify groups of wild strains that are probably of a common origin, most likely like the local microbiota, as similarly observed by Kishkovskaia et al (2017).…”

“…Understanding native yeast population and evolution dynamics is important in order to comprehend how the stressing factors of the fermentation process impact the commercial yeast strains go through and how they interact with competitive invasive strains (Payen et al, 2014). Additionally, repetitive DNA sequences may display inter-specific patterns, punctual variations in position and number of such regions constitutes a genetic fingerprinting that permits to identify and differentiate yeast strains or clonal variants from a given local microbiota (Xufre et al, 2011).…”