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      Utilização do Intron Splice Site primer EI-1 na discriminação de leveduras contaminantes do processo de fermentação alcoólica Translated title: Use of Intron Splice Site primer EI-1 in the discrimination of contaminant yeasts from the alcoholic fermentation process

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          Abstract

          Neste trabalho, utilizamos o Intron Splice Site primer EI-1 para a análise do perfil de amplificação de diferentes espécies de leveduras consideradas contaminantes no processo de fermentação alcoólica, originadas de uma destilaria no Estado da Paraíba na safra 2004/2005. Foram realizadas as etapas analíticas para discriminação molecular das leveduras a partir da extração do DNA total, amplificação por PCR e análise do perfil genético. Os resultados obtidos indicam que o Intron Splice Site primer EI-1é muito eficaz na discriminação das diferentes espécies de Saccharomyces e não Saccharomyces, evidenciando padrões de bandas específicos para as leveduras analisadas. Este primer, por ser complementar a uma região muito conservada do genoma das leveduras, mostrou-se incapaz de uma discriminação intraespecífica. Isto demonstra a utilidade deste marcador no auxílio à taxonomia de leveduras.

          Translated abstract

          In the present study, the Intron Splice Site primer EI-1 was used for the analysis of the profile amplification of different species of yeasts considered contaminants (16) in the alcoholic fermentation process obtained from a distillery in the State of Paraíba in 2004/2005. Analytic stages for the molecular discrimination of the yeasts from the total DNA extraction, PCR amplification, and genetic profile analysis were performed. The results indicates that Intron Splice Site primer EI-1 is highly efficient at discriminating between different Saccharomyces and non-Saccharomyces species showing specific band standards for the yeasts analyzed. This primer, which is part of a conserved region of yeast genome, proved incapable of intra-specific discrimination. This finding demonstrates that this marker may be useful in yeast taxonomy.

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          Most cited references15

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          Rapid detection and quantification of yeast species during spontaneous wine fermentation by PCR-RFLP analysis of the rDNA ITS region.

          PCR-RFLP analysis of the rDNA-ITS (internal transcribed spacer) region was applied to 174 yeast strains belonging to 30 species of oenological significance and including 27 type strains in order to define a rapid identification protocol for yeast colonies. DraI-or HaeIII-PCR-RFLP patterns were species-specific with the exception of teleomorphic and anamorphic forms. An improved protocol taking about 30 h was used for the detection and quantification of yeast species occurring in the course of a spontaneous wine fermentation at industrial level. Wine samples were taken and plated daily on an agar medium and the developed colonies were analysed by PCR-RFLP after 24 h of incubation. A representative sample of these colonies was also identified by traditional methods. Both procedures gave identical results. However, PCR-RFLP analysis allowed a more precise enumeration of the yeast populations, proving to be a reliable and simple method for monitoring the development of the yeast community throughout wine fermentation.
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            Yeast communities and genetic polymorphism of Saccharomyces cerevisiae strains associated with artisanal fermentation in Brazil.

            Yeast communities and genetic polymorphism of prevalent Saccharomyces cerevisiae strains isolated from the spontaneous fermentation of the sugarcane juice during the production of aguardente in three distilleries in the state of Minas Gerais, Brazil, were studied. S. cerevisiae was the prevalent species during the process of aguardente production, but Schizosaccharomyces pombe was predominant in old fermentations in one distillery. Transient yeast species were found in a variable number, probably due to the daily addition of sugarcane juice, and they were different for each of the three distilleries studied. PFGE and PCR analysis of the predominant strains of S. cerevisiae isolated from the fermented must showed a high degree of genetic polymorphism among the three distilleries. A high molecular variability of S. cerevisae strains was also observed among strains isolated from the same vat at different fermentation ages. Our results showed that there was a succession of geneticly different strains of S. cerevisae during the process of aguardente production.
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              Genetic diversity of Saccharomyces cerevisiae strains during the 24 h fermentative cycle for the production of the artisanal Brazilian cachaça.

              Characterization of yeast populations and genetic polymorphism of Saccharomyces cerevisiae strains collected during the short fermentative cycles from the spontaneous fermentations during the artisanal cachaça production. The prevalent S. cerevisiae strains were analysed by PFG and RAPD-PCR using primers EI1 and M13. The molecular analysis have showed a high degree of genetic polymorphism among the strains within a 24 h fermentative cycle. The genetic diversity observed in the S. cerevisiae strains may be occurring due to the existence of a large number of individual genotypes within the species. The unique characteristics of the cachaça fermentation process probably allows for a faster detection of molecular polymorphisms of yeast strains than other types of fermentations. Spontaneous fermentations to produce cachaça, due to their characteristics, are an excellent model for the study of molecular diversity of S. cerevisiae strains during the production of fermented beverages.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Journal
                cta
                Food Science and Technology (Campinas)
                Food Sci. Technol (Campinas)
                Sociedade Brasileira de Ciência e Tecnologia de Alimentos (Campinas )
                1678-457X
                September 2010
                : 30
                : 3
                : 761-765
                Affiliations
                [1 ] Universidade Federal de Pernambuco Brazil
                [2 ] Universidade Federal de Pernambuco Brazil
                [3 ] Universidade Federal de Pernambuco Brazil
                [4 ] Universidade Federal de Pernambuco Brazil
                Article
                S0101-20612010000300030
                10.1590/S0101-20612010000300030
                e094a72f-563f-486c-8b2a-81238634c423

                http://creativecommons.org/licenses/by/4.0/

                History
                Product

                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=0101-2061&lng=en
                Categories
                FOOD SCIENCE & TECHNOLOGY

                Food science & Technology
                contaminant yeasts,Saccharomyces cerevisiae,Intron,leveduras contaminantes

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