Genome Sequence of the Native Apiculate Wine Yeast Hanseniaspora vineae T02/19AF

The use of novel yeast strains for winemaking improves quality and provides variety including subtle characteristic differences in fine wines. Here we report the first genome of a yeast strain native to Uruguay, Hanseniaspora vineae T02/19AF, which has been shown to positively contribute to aroma and wine quality.Fil: Giorello, Facundo M.. Universidad de la República; UruguayFil: Berná, Luisa. Instituto Pasteur de Montevideo; UruguayFil: Greif, Gonzalo. Instituto Pasteur de Montevideo; UruguayFil: Camesasca, Laura. Inst. de Investigaciones Biológicas Clemente Estable; UruguayFil: Salzman, Valentina. Instituto Pasteur de Montevideo; Uruguay. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Medina, Karina. Universidad de la Republica. Facultad de Química; UruguayFil: Robello, Carlos. Instituto Pasteur de Montevideo; UruguayFil: Gaggero, Carina. Inst. de Investigaciones Biológicas Clemente Estable; UruguayFil: Aguilar, Pablo S.. Instituto Pasteur de Montevideo; UruguayFil: Carrau, Francisco. Sección Enología; Urugua

The Physcomitrella patens unique alpha-dioxygenase participates in both developmental processes and defense responses

[Background] Plant α-dioxygenases catalyze the incorporation of molecular oxygen into polyunsaturated fatty acids leading to the formation of oxylipins. In flowering plants, two main groups of α-DOXs have been described. While the α-DOX1 isoforms are mainly involved in defense responses against microbial infection and herbivores, the α-DOX2 isoforms are mostly related to development. To gain insight into the roles played by these enzymes during land plant evolution, we performed biochemical, genetic and molecular analyses to examine the function of the single copy moss Physcomitrella patens α-DOX (Ppα-DOX) in development and defense against pathogens.[Results] Recombinant Ppα-DOX protein catalyzed the conversion of fatty acids into 2-hydroperoxy derivatives with a substrate preference for α-linolenic, linoleic and palmitic acids. Ppα-DOX is expressed during development in tips of young protonemal filaments with maximum expression levels in mitotically active undifferentiated apical cells. In leafy gametophores, Ppα-DOX is expressed in auxin producing tissues, including rhizoid and axillary hairs. Ppα-DOX transcript levels and Ppα-DOX activity increased in moss tissues infected with Botrytis cinerea or treated with Pectobacterium carotovorum elicitors. In B. cinerea infected leaves, Ppα-DOX-GUS proteins accumulated in cells surrounding infected cells, suggesting a protective mechanism. Targeted disruption of Ppα-DOX did not cause a visible developmental alteration and did not compromise the defense response. However, overexpressing Ppα-DOX, or incubating wild-type tissues with Ppα-DOX-derived oxylipins, principally the aldehyde heptadecatrienal, resulted in smaller moss colonies with less protonemal tissues, due to a reduction of caulonemal filament growth and a reduction of chloronemal cell size compared with normal tissues. In addition, Ppα-DOX overexpression and treatments with Ppα-DOX-derived oxylipins reduced cellular damage caused by elicitors of P. carotovorum.[Conclusions] Our study shows that the unique α-DOX of the primitive land plant P. patens, although apparently not crucial, participates both in development and in the defense response against pathogens, suggesting that α-DOXs from flowering plants could have originated by duplication and successive functional diversification after the divergence from bryophytes.This work was supported by Agencia Nacional de Investigación e Innovación (ANII) [grants FCE2007_376, FCE2011_6095, fellowships BE_POS_2009_726 (A. Castro) and BE_POS_2010_2533 (L. Machado)], UdelaR Uruguay/CSIC Spain (Joint project), the Swedish Research Council, and Programa de Desarrollo de las Ciencias Básicas (PEDECIBA) Uruguay. The Ppα-DOX cDNA was obtained from the RIKEN Biological Research Center, Tsukuba, Japan

Yeast diversity and native vigor for flavor phenotypes

Saccharomyces cerevisiae, the yeast used widely for beer, bread, cider, and wine production, is the most resourceful eukaryotic model used for genetic engineering. A typical concern about using engineered yeasts for food production might be negative consumer perception of genetically modified organisms. However, we believe the true pitfall of using genetically modified yeasts is their limited capacity to either refine or improve the sensory properties of fermented foods under real production conditions. Alternatively, yeast diversity screening to improve the aroma and flavors could offer groundbreaking opportunities in food biotechnology. We propose a ‘Yeast Flavor Diversity Screening’ strategy which integrates knowledge from sensory analysis and natural whole-genome evolution with information about flavor metabolic networks and their regulation.Fil: Carrau, Francisco. Universidad de la República; UruguayFil: Gaggero, Carina. Instituto de Investigaciones Biológicas ; UruguayFil: Aguilar, Pablo Sebastián. Instituto Pasteur de Montevideo; Uruguay. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Molecular Relationships Between Saccharomyces cerevisiae Strains Involved in Winemaking from Mendoza, Argentina

Three molecular typing techniques were applied to assess the molecular relationships of Saccharomyces cerevisiae strains isolated from winery equipment, grapes, and spontaneous fermentation in a cellar located in ‘‘Zona Alta del Rı´o Mendoza’’ (Argentina). In addition, commercial Saccharomyces strains widely used in this region were also included. Interdelta PCR typing, mtDNA restriction analysis, and microsatellite (SSR) genotyping were applied. Dendrograms were constructed based on similarity among different patterns of bands. The combination of the three techniques discriminated 34 strains among the 35 isolates. The results of this study show the complex relationships found at molecular level among the isolates that share the same ecological environment, i.e., the winemaking process. With a few exceptions, the yeast isolates were generally clustered in different ways, depending on the typing technique employed. Three clusters were conserved independently of the molecular method applied. These groups of yeasts always clustered together and had high degree of similarity. Furthermore, the dendrograms mostly showed clusters combining strains from winery and fermentation simultaneously. Most of the commercial strains included in this study were clustered separately from the other isolates analyzed, and just a few of them grouped with the strains mainly isolated from spontaneous fermentation. Only one commercial strain was clustered repetitively with a noncommercial strain isolated from spontaneous fermentation in the three dendrograms. On the other hand, this study has demonstrated the importance of selecting an appropriate molecular method according to the main objectives of the research.Fil: Mercado, Laura Analia. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Mendoza-San Juan. Estación Experimental Agropecuaria Mendoza. Centro de Estudios Enológicos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; ArgentinaFil: Jubany, Sandra. Instituto de Investigaciones Biológicas "Clemente Estable"; UruguayFil: Gaggero, Carina. Instituto de Investigaciones Biológicas "Clemente Estable"; UruguayFil: Masuelli, Ricardo Williams. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Mendoza-San Juan. Estación Experimental Agropecuaria La Consulta; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias; ArgentinaFil: Combina, Mariana. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Mendoza-San Juan. Estación Experimental Agropecuaria La Consulta; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentin

De novo synthesis of benzenoid compounds by the yeast hanseniaspora vineae increases the flavor diversity of wines

Benzyl alcohol and other benzenoid-derived metabolites of particular importance in plants confer floral and fruity flavors to wines. Among the volatile aroma components in Vitis vinifera grape varieties, benzyl alcohol is present in its free and glycosylated forms. These compounds are considered to originate from grapes only and not from fermentative processes. We have found increased levels of benzyl alcohol in red Tannat wine compared to that in grape juice, suggesting de novo formation of this metabolite during vinification. In this work, we show that benzyl alcohol, benzaldehyde, p-hydroxybenzaldehyde, and p-hydroxybenzyl alcohol are synthesized de novo in the absence of grape-derived precursors by Hanseniaspora vineae. Levels of benzyl alcohol produced by 11 different H. vineae strains were 20-200 times higher than those measured in fermentations with Saccharomyces cerevisiae strains. These results show that H. vineae contributes to flavor diversity by increasing grape variety aroma concentration in a chemically defined medium. Feeding experiments with phenylalanine, tryptophan, tyrosine, p-aminobenzoic acid, and ammonium in an artificial medium were tested to evaluate the effect of these compounds either as precursors or as potential pathway regulators for the formation of benzenoid-derived aromas. Genomic analysis shows that the phenylalanine ammonia-lyase (PAL) and tyrosine ammonia lyase (TAL) pathways, used by plants to generate benzyl alcohols from aromatic amino acids, are absent in the H. vineae genome. Consequently, alternative pathways derived from chorismate with mandelate as an intermediate are discussed.Fil: Martín, Valentina. Universidad de la República; Uruguay. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Giorello, Facundo. Universidad de la República; UruguayFil: Fariña, Laura. Universidad de la República; Uruguay. Instituto de Investigaciones Biológicas "Clemente Estable"; UruguayFil: Minteguiaga, Manuel. Universidad de la República; UruguayFil: Salzman, Valentina. Instituto Pasteur de Montevideo; Uruguay. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Boido, Eduardo. Universidad de la República; UruguayFil: Aguilar, Pablo Sebastián. Instituto Pasteur de Montevideo; Uruguay. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Gaggero, Carina. Instituto de Investigaciones Biológicas "Clemente Estable"; UruguayFil: Dellacassa, Eduardo. Universidad de la República; UruguayFil: Mas, Albert. Universitat Rovira I Virgili; EspañaFil: Carrau, Francisco. Universidad de la República; Urugua

Vitis vinifera Tannat, the result of a multidisciplinary research

Tema del mesLa calidad de la uva, y por tanto del vino, es el resultado de la interacción de numerosos factores que involucran aspectos biológicos (variedad, clon, portainjerto, estado sanitario), físicos (tipo de suelo y su manejo), climáticos (temperatura, pluviometría, luz) y culturales (densidad de plantación, conducción, poda, carga de fruta, manejo de la vegetación, fertilización). La conjunción y el manejo armónico de estos factores condicionarán las potencialidades de un viñedo y determinará la cantidad de fruta capaz de soportar la planta. Los viticultores uruguayos han buscado incidir en estos aspectos reemplazando las viejas plantas Tannat por nuevos clones comerciales de esta variedad. Para intentar evaluar el impacto de esta decisión, nuestro grupo evaluó las relaciones genéticas entre los clones Tannat antiguos, presentes en Uruguay, y los clones comerciales franceses, introducidos recientemente. Los resultados demostraron que los clones analizados representan un pool genéticamente uniforme, donde se pueden discriminar dos grupos mediante marcadores moleculares y por la composición de componentes volátiles unidos a azúcares (glicósidos). Este aspecto es desafiante, ya que una de las características económicas más significativas de cultivar es el aroma que la uva puede impartirle al vino, ya que los constituyentes volátiles de la uva son responsables de los principales aromas del mosto y proveen las bases del carácter varietal. La calidad de un vino también es consecuencia de los procesos de fermentación (alcohólica y maloláctica). La fermentación aumenta la complejidad aromática del vino, ayudando en la extracción de los sólidos del mosto, modificando compuestos derivados de la uva y produciendo una importante cantidad de compuestos aromáticos (alcoholes, ésteres, ácidos, aldehídos). Aunque estos compuestos pueden encontrarse en muchos vinos, el tipo de proceso ejecutado puede causar cambios importantes en sus concentraciones. Para el vino Tannat esto se evaluó estudiando el perfil aromático por análisis sensorial y por análisis químico, utilizando una separación cromatográfica con jueces capaces de percibir el aroma de los componentes separados (olfatomería). Los vinos tintos Tannat también poseen contenidos elevados de taninos y un color intenso, características responsables de la originalidad de estos vinos. Las correlaciones entre las diferentes familias de pigmentos estudiados por técnicas separativas y espectroscópicas y parámetros que definen las notas de color (CIELAB), revelaron la importancia de las variaciones encontradas en antocianos, flavanol-antocianos y piranoantocianinas en el color y palatabilidad de estos vinos. El enfoque presentado muestra la integración multidisciplinaria que define nuestro grupo, que busca aportar herramientas objetivas para aplicar el concepto genérico de “Enología de Mínima Intervención” a la producción de vinos Tannat.The quality of the grapes, and thus the wine, is the result of the interaction of many factors involving biological (variety, clone, rootstock and sanitary status), physical (soil type and its management), climate (temperature, rainfall, light) and cultural (planting density, canopy management, ripening, fruit load, fertilization). The effect produced by the sum of these factors, and their control, will condition the potential of a vineyard determining the amount of fruit capable of supporting the plant. The Uruguayan winemakers have intended to influence on this situation for the Vitis vinifera cv Tannat, replacing the old plants by new commercially available clones of this variety. In order to assess the impact of this decision, our research group studied the genetic relationships among the old Tannat clones present in Uruguay, and the French commercial ones recently introduced. The results demonstrated that the clones evaluated represent a genetically uniform pool, where two groups can be discriminated using both molecular markers and the composition of glicosylated aroma compounds. This aspect has a particular relevance as the scent that the grape can impart to the wine represents one of the most significant economic characteristics of a cultivar. Secondary metabolites from grapes are responsible for the main must aromatic constituents providing the basis of the varietal character. The wine quality is also a result of the fermentation processes (alcoholic and malolactic). The fermentations increase the wine aromatic complexity, assisting the removal of solids from the must and produce chemical modifications on those compounds derived from grapes producing a significant amount of metabolites (alcohols, esters, acids, aldehydes). Although these compounds can be found in many wines, the kind of process executed can cause major changes in their concentrations. These aspects were studied on Tannat wines studying their flavor profile using different approaches: descriptive sensory analysis, chemical analysis and gas chromatographyolfactometry. Tannin contents and the intense color of Tannat red wines are also features responsible for the originality of these wines. The correlations between the different pigment families studied by separation and spectroscopic tools and CIELAB parameters revealed the importance of the variations found in anthocyanins, flavanol-anthocyanins, pyranoanthocyanins fractions and the color and palatability of these wines. The results above presented shows the multidisciplinary integration that defines our group, which seeks to provide objective tools to implement the generic concept of “Minimum Intervention Oenology” to produce Tannat wines