Differentiation of species of the genus Saccharomyces using biomolecular fingerprinting methods

Abstract

The genus Saccharomyces comprises very closely related species. This high degree of relationship makes a simple identification and differentiation of strains difficult since these species are hardly discriminable by their morphological and physiological features. A sequence analysis of ribosomal DNA and the corresponding internal transcribed spacers can only rarely be successfully applied. In this study, we proved the applicability of a novel DNA fingerprinting method, the SAPD-PCR (specifically amplified polymorphic DNA) and of MALDI-TOF-MS (matrix-assisted laser desorption ionization time-of-flight mass spectrometry) fingerprinting with the MALDI Biotyper for the differentiation of species belonging to the genus Saccharomyces. It was possible with SAPD-PCR to create specific banding patterns for all Saccharomyces species. Different strains of the same species produced nearly the same banding patterns. Specific and reproducible reference spectra could be generated for each of the strains with the MALDI Biotyper. Therefore, SAPD-PCR and MALDI-TOF-MS can be fast and reliable tools to identify these related Saccharomyces species which are applied in many biotechnological processes.

MALDI-TOF mass spectrometry fingerprinting: A diagnostic tool to differentiate dematiaceous fungi Stachybotrys chartarum and Stachybotrys chlorohalonata

“…Samples were analyzed with a Microflex LT-mass spectrometer (Bruker Daltonik GmbH, Bremen, Germany). Calibration of mass spectra and conditions for acquiring mass spectra and identifying fungal samples are described in Blaettel et al (2013). Used software version was MALDI Biotyper 2.0.…”

MALDI-TOF mass spectroscopy of yeasts and filamentous fungi for research and diagnostics in the agricultural value chain

“…In the case of pathogenic yeasts, the sibling species of the Cryptococcus gattii/Cryptococcus neoformans complex, which cannot be discriminated by routine biochemical techniques, have been distinguished by MALDI biotyping [71]. [72]. Finally, MALDI-TOF MS more rapidly identified reduced susceptibility to caspofungin or triazoles in Candida and Aspergillus species, as compared to classical determination of the minimum inhibitory concentration [39,73,74].…”

“…These studies document that in particular the genera Fusarium, Trichoderma, and Saccharomyces are being used as models for the development of MALDI-TOF MS applications and for assessing the potential of this technology. In contrast to clinical studies, naturally [127] occurring yeasts have been rarely studied by MALDI-TOF MS, except in the context of fermentations, for example, during winemaking and brewing [19,72,87,88]. In general, the large majority of (non-clinical) fungal MALDI-TOF MS studies were directed towards phytopathogens (e.g., Aspergillus, Fusarium, Monilinia, Penicillium, Puccinia, mildews) or potential biocontrol strains (e.g., Trichoderma, Metarhizium).…”

An Exemplary Model Study for Overcoming Stuck Fermentation during Spontaneous Fermentation with the Aid of a Saccharomyces Triple Hybrid

“…The DNA fingerprinting method SAPD-PCR was carried out according to Blättel et al (2013) for the further identification of the Saccharomyces species.…”

“…The most important and well-known natural yeast in spontaneous fermentations is Saccharomyces cerevisiae, although Saccharomyces bayanus may also be found in the later stages of fermentations (Ultee et al, 2013). However, hybrids of Saccharomyces species have also been identified as being involved in the fermentation process during winemaking (González et al, 2006;Arroyo-López et al, 2009;Blättel et al, 2013). González et al (2006) described the hybrid S. cerevisiae × S. kudriavzevii which could be advantageous for specific fermentation conditions due to the combined characteristics of both parents.…”