ABSTRACT
Genomic studies of yeasts from the wild have increased considerably in the past few years. This revolution has been fueled by advances in high-throughput sequencing technologies and a better understanding of yeast ecology and phylogeography, especially for biotechnologically important species. The present review aims to first introduce new bioinformatic tools available for the generation and analysis of yeast genomes. We also assess the accumulated genomic data of wild isolates of industrially relevant species, such as Saccharomyces spp., which provide unique opportunities to further investigate the domestication processes associated with the fermentation industry and opportunistic pathogenesis. The availability of genome sequences of other less conventional yeasts obtained from the wild has also increased substantially, including representatives of the phyla Ascomycota (e.g. Hanseniaspora) and Basidiomycota (e.g. Phaffia). Here, we review salient examples of both fundamental and applied research that demonstrate the importance of continuing to sequence and analyze genomes of wild yeasts.
Subject(s)
Computational Biology/methods , Fermentation , Genome, Fungal , Saccharomyces/genetics , Yeasts/genetics , Ascomycota/genetics , Basidiomycota/genetics , Chromosome Mapping , Environment , Genomics , Hanseniaspora/genetics , High-Throughput Nucleotide Sequencing , Wine/analysis , Yeasts/classificationABSTRACT
UNLABELLED: We investigated the presence of the yeast Dekkera bruxellensis in samples collected at three points surrounding the industrial alcoholic fermentation plants of two distilleries where there are often cases of contamination caused by this yeast: this involved sugar cane wash water, feeding sugar cane juice and vinasse from the treatment pond. Total yeast was isolated in WLN medium with bromocresol green and cycloheximide and further selected on the basis of its ability to grow in synthetic medium containing nitrate. Following this, colonies were selected from the distribution on nitrate plates and identified by amplification with species-specific primers and DNA sequencing of the 26S-D1/D2 locus. The results showed that D. bruxellensis is introduced through the feeding substrate, which suggests that its cells originated with the harvested cane. Subsequently, its population circulates as a result of the reuse of water for washing the cane, in a continuous re-inoculation of the plant with yeasts. Furthermore, the yeast population is formed in the vinasse by the addition of wash water into the treatment ponds and then reintroduced to the culture fields by fertigation, so that the process can be renewed in the following season. It is now possible to adopt sanitation procedures that can prevent the entry of the contamination to the fermentation process. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of the yeast Dekkera bruxellensis is sometimes attributed to a decline in the industrial productivity of ethanol since it has a more limited fermentation capacity than Saccharomyces cerevisiae. Although its adaptability to the industrial environment has been noted, so far, there has been no evidence to determine the source of this contamination. In this study, we provide evidence to show that D. bruxellensis comes from the fields together with the harvested cane and is then accumulated and recirculated. It might be possible to prevent the accumulation of this yeast by carrying out sanitation controls during the harvesting season.
Subject(s)
Bioreactors/microbiology , Dekkera/growth & development , Dekkera/metabolism , Ethanol/metabolism , Saccharum/microbiology , Dekkera/genetics , Fermentation/physiology , Industrial Microbiology/methods , Nitrates , Saccharomyces cerevisiae/metabolism , Water MicrobiologyABSTRACT
The number of killer, neutral and sensitive yeasts was determined from strains isolated from substrates related to alcoholic fermentations. From 113 isolates, 24 showed killer activity against NCYC 1006 (standard sensitive strain), while 30 were sensitive to NCYC 738 (standard killer strain), and 59 had no reaction in assays at 25-27°C. Two wild yeast strains of Saccharomyces cerevisiae and one of Candida colliculosa were tested against 10 standard killer strains and one standard sensitive strain in a cell x cell and well-test assays at four different pHs. None of the isolates displayed strong killer activity or were sensitive to the standard strains. All belonged to the neutral type. It was concluded that although the number of killer strains was high, this character cannot be used to protect ethanol fermentation processes against yeast contaminants like those which form cell clusters.
Avaliou-se o número de linhagens 'killer', sensíveis e neutras em leveduras isoladas de substratos relacionados à fermentação etanólica. Das 113 linhagens, 24 mostraram atividade 'Killer' contra o isolado NCYC 1006 (padrão de sensibilidade), 30 foram sensíveis ao isolado NCYC 738 (padrão 'Killer') e 59 apresentaram reação neutra para ambos os isolados. Três cepas de leveduras selvagens do processo (duas de Sacch. cerevisiae e uma de Candida colliculosa, que formam cachos de células não separáveis por tratamentos químicos ou físicos), foram testadas contra 10 isolados padrões do tipo 'Killer' e um isolado padrão de sensibilidade. Os ensaios células x células e células X toxinas foram realizados em diferentes pH e a 30ºC. As três cepas contaminantes mostraram reação neutra a todos os isolados padrões testados. Apesar do alto número de linhagens 'Killer' entre aquelas testadas, concluiu-se que este caráter não pode ser utilizado na proteção do processo de fermentação etanólica contra essas leveduras selvagens contaminantes.
ABSTRACT
The number of killer, neutral and sensitive yeasts was determined from strains isolated from substrates related to alcoholic fermentations. From 113 isolates, 24 showed killer activity against NCYC 1006 (standard sensitive strain), while 30 were sensitive to NCYC 738 (standard killer strain), and 59 had no reaction in assays at 25-27°C. Two wild yeast strains of Saccharomyces cerevisiae and one of Candida colliculosa were tested against 10 standard killer strains and one standard sensitive strain in a cell x cell and well-test assays at four different pHs. None of the isolates displayed strong killer activity or were sensitive to the standard strains. All belonged to the neutral type. It was concluded that although the number of killer strains was high, this character cannot be used to protect ethanol fermentation processes against yeast contaminants like those which form cell clusters.
Avaliou-se o número de linhagens 'killer', sensíveis e neutras em leveduras isoladas de substratos relacionados à fermentação etanólica. Das 113 linhagens, 24 mostraram atividade 'Killer' contra o isolado NCYC 1006 (padrão de sensibilidade), 30 foram sensíveis ao isolado NCYC 738 (padrão 'Killer') e 59 apresentaram reação neutra para ambos os isolados. Três cepas de leveduras selvagens do processo (duas de Sacch. cerevisiae e uma de Candida colliculosa, que formam cachos de células não separáveis por tratamentos químicos ou físicos), foram testadas contra 10 isolados padrões do tipo 'Killer' e um isolado padrão de sensibilidade. Os ensaios células x células e células X toxinas foram realizados em diferentes pH e a 30ºC. As três cepas contaminantes mostraram reação neutra a todos os isolados padrões testados. Apesar do alto número de linhagens 'Killer' entre aquelas testadas, concluiu-se que este caráter não pode ser utilizado na proteção do processo de fermentação etanólica contra essas leveduras selvagens contaminantes.