Kinetics and mathematical model of killer/sensitive interaction under different physicochemical conditions of must/wine: a study from a biological point of view.

Fermentation of grape must to wine is carried out by a complex microbial mixture, which also involves spoilage yeasts of wine. The latter yeasts produce organoleptic changes that cause significant economic losses to the wine industry. SO2 is traditionally used to control this spoilage populations, but because of its harmful effects on human health, biocontrol has emerged as an alternative treatment. Although studies have been carried out to select biocontroller yeasts and examine their underlying mechanisms of action, reports on their application have not been published yet. To better understand the interaction and the successful application of biocontrol, the use of mathematical models, among other methods, is important, as they facilitate the prediction of success or failure of the antagonist. The objective of the present study was to use an existing mathematical model to obtain information about the yeast's interaction assayed and to validate its predictive use under different physicochemical conditions during the wine fermentation, and eventually predict biocontrol kinetics. The mathematical model was applied to the fermentation conditions and provided information on the kinetic parameters of the biocontrol interaction and allowed interpretations about other parameters. The model was applied in the different physicochemical conditions for the biocontrol and did not fit correctly to experimental data, and therefore an improvement was proposed which was successful and presented new hypotheses.

[Behaviour of a wild Saccharomyces cerevisae killer yeast and its isogenic sensitive one with respect to different nitrogen sources in mixed cultures]. / Comportamiento de una cepa salvaje de saccharomyces cerevisiae killer y su isogénica sensible respecto de diferentes fuentes de nitrógeno en cultivos mixtos.

Wine chemical composition is the outcome of complex chemosensory interactions that are difficult to predict because of the influences of many variables, like as yeast-yeast interactions. Killer phenomenon implicates the secretion of a toxic protein by some yeasts, that kill other yeasts called sensitive. The knowledge of the behaviour of killer-sensitive mixed cultures in aerobic conditions is useful to be related with the first stages of oenological fermentation. In these stages it can be defined the killer prevalence in the medium. Also, the use of cured plasmid mutants allows better comparisons. The objective was to analyse the mechanism of substrate competition in Saccharomyces cerevisiae killer strains and its sensitive cured plasmid mutant, using different nitrogen sources. When samples were incubated at the toxin inactivation temperature, the infraestimation of sensitive cells is avoided. Results obtained in co-cultures (50%K-50%S) show the role of the nitrogen source in killer activity. Results obtained with 10%K-90%S inoculum, show that there are another competence variables than the competitive exclusion model for killer yeasts.

[Yeast diversity in Bulnesia retama and Larrea divaricata canopies and associated soils]. / Diversidad de levaduras en canopias y suelos asociados con Bulnesia retama y Larrea divaricata.

Bush like vegetation dominates arid environments, and there is nutrients accumulation under shrub canopies and relatively unfertile soils between vegetal patches areas. Plants are one of the most common habitats for yeasts. There are many reports about yeasts inhabiting different plant components. Nevertheless, there are no reports about yeasts associated with Zygophyllaceae, an important shrub family of the Argentinean Province of Monte. The objective of this work was to analyzed yeast biodiversity of Bulnesia retama and Larrea divaricata canopies and associated soils, at Medanos Grandes of Caucete, San Juan, Argentina. Eighty seven (87) isolated yeasts were identified. From B. retama canopy and associated soil was observed a larger taxonomical diversity respect to L. divaricata. Nine (9) and ten (10) species were isolated from canopy and associated soil of B. retama, respectively. From L. divaricata canopy were 4 species and 3 species from its associated soil isolated. Identified genera were: Candida, Debaryomyces, Dekkera, Saccharomyces, Torulaspora, Sporidiobolus and Pichia. Fourteen (14) species were found at all microenvironments.

Antifungal modes of action of Saccharomyces and other biocontrol yeasts against fungi isolated from sour and grey rots.

The aim of this study was to determine the putative modes of action of 59 viticultural yeasts (31 Saccharomyces and 28 non-Saccharomyces) that inhibited fungi isolated from sour and grey rot in grapes. Inhibition of fungal mycelial growth by metabolites, enzyme activities (laminarinases, chitinases), antifungal volatiles, competition for nutrients (siderophores, Niche Overlap Index (NOI)), inhibition of fungal spore germination and decreased germinal tube length and induction of resistance were assayed. Biofungicide yeasts were classified into "antifungal patterns", according to their mechanisms of action. Thirty isolates presented at least two of the mechanisms assayed. We propose that inhibition of fungal mycelial growth by metabolites, laminarinases, competition for nutrients, inhibition of fungal spore germination and decreased germinal tube length, and antifungal volatiles by Saccharomyces and non-Saccharomyces viticultural yeasts is used as putative biocontrol mechanisms against phytopathogenic fungi. Twenty-four different antifungal patterns were identified. Siderophore production (N)and a combination of siderophore production and NOI>0.92 (M)were the most frequent antifungal patterns observed in the biofungicide yeasts assayed. Elucidation of these mechanisms could be useful for optimization of an inoculum formulation, resulting in a more consistent control of grey and sour rot with Saccharomyces and non-Saccharomyces biocontrol yeasts.

Comportamiento de una cepa salvaje de Saccharomyces cerevisiae killer y su isogénica sensible respecto de diferentes fuentes de nitrógeno en cultivos mixtos / Behaviour of a wild Saccharomyces cerevisiae killer yeast and its isogenic sensitive one with respect to different nitrogen sources in mixed cultures

La composición química del vino constituye el fundamento de la posterior respuesta sensorial del producto y está determinada por varios factores, como las relaciones levadura-levadura. Se denomina fenómeno killer a la secreción por parte de ciertas cepas de levadura de una proteína tóxica que mata a células denominadas sensibles. El conocimiento del comportamiento en condición aeróbica de cultivos mixtos killer-sensible es útil para relacionarlo con la primera fase de la fermentación enológica, ya que en ella puede definirse la prevalencia o no de la cepa killer. Además, el empleo de mutantes con el plásmido curado permite comparaciones más precisas. El objetivo fue analizar el mecanismo de competencia por sustrato en levaduras killer de Saccharomyces cerevisiae y su mutante sensible con el plásmido curado, empleando distintas fuentes de nitrógeno. Si las muestras se incuban a temperatura de inactivación de la toxina, se evita la infraestimación de células sensibles. Los resultados del co-cultivo de las cepas en proporciones iguales muestran el rol desempeñado por la fuente de nitrógeno en la actividad killer. Cuando el inóculo es 10%K-90%S, el modelo de exclusión competitiva planteado para levaduras killer deja paso a otras variables de competencia.

Wine chemical composition is the outcome of complex chemosensory interactions that are difficult to predict because of the influences of many variables, like as yeast-yeast interactions. Killer phenomenon implicates the secretion of a toxic protein by some yeasts, that kill other yeasts called sensitive. The knowledge of the behaviour of killer-sensitive mixed cultures in aerobic conditions is useful to be related with the first stages of oenological fermentation. In these stages it can be defined the killer prevalence in the medium. Also, the use of cured plasmid mutants allows better comparisons. The objective was to analyse the mechanism of substrate competition in Saccharomyces cerevisiae killer strains and its sensitive cured plasmid mutant, using different nitrogen sources. When samples were incubated at the toxin inactivation temperature, the infraestimation of sensitive cells is avoided. Results obtained in co-cultures (50%K-50%S) show the role of the nitrogen source in killer activity. Results obtained with 10%K-90%S inoculum, show that there are another competence variables than the competitive exclusion model for killer yeasts.

Diversidad de levaduras en canopias y suelos asociados con Bulnesia retama y Larrea divaricata / Yeast diversity in Bulnesia retama and Larrea divaricata canopies and associated soils.

Los ambientes áridos están dominados por vegetación arbustiva, con acumulación de nutrientes bajo la canopia de los arbustos y con suelos relativamente infértiles en los interparches. Los distintos componentes de los vegetales constituyen uno de los hábitat más comunes para las levaduras. Existen numerosos antecedentes acerca de investigaciones sobre levaduras cuyo hábitat lo constituyen árboles y arbustos, sin embargo no existen referencias sobre levaduras asociadas a las Zigofiláceas, una familia de matorrales preponderantes en la Provincia de Monte argentino. El objetivo de este trabajo fue conocer la biodiversidad de levaduras en suelos y canopias asociados con Bulnesia retama y Larrea divaricata, en los Médanos Grandes de Caucete, San Juan, Argentina. Sobre un total de 87 aislamientos de levaduras identificados, se observó una mayor diversidad taxonómica en las asociadas tanto al suelo como a la parte aérea de B. retama, respecto de L. divaricata. A partir de la canopia de B. retama y su suelo asociado se aislaron 9 y 10 especies de levaduras respectivamente, mientras que de la parte aérea y suelo de L. divaricata 4 y 3. Los géneros identificados fueron: Candida, Debaryomyces, Dekkera, Saccharomyces, Torulaspora, Sporidiobolus y Pichia. En total se encontraron 14 especies en todos los microambientes.

Bush like vegetation dominates arid environments, and there is nutrients accumulation under shrub canopies and relatively unfertile soils between vegetal patches areas. Plants are one of the most common habitats for yeasts. There are many reports about yeasts inhabiting different plant components. Nevertheless, there are no reports about yeasts associated with Zigophyllaceae, an important shrub family of the Argentinean Province of Monte. The objective of this work was to analyzed yeast biodiversity of Bulnesia retama and Larrea divaricata canopies and associated soils, at Médanos Grandes of Caucete, San Juan, Argentina. Eighty seven (87) isolated yeasts were identified. From B. retama canopy and associated soil was observed a larger taxonomical diversity respect to L. divaricata. Nine (9) and ten (10) species were isolated from canopy and associated soil of B. retama, respectively. From L. divaricata canopy were 4 species and 3 species from its associated soil isolated. Identified genera were: Candida, Debaryomyces, Dekkera, Saccharomyces, Torulaspora, Sporidiobolus and Pichia. Fourteen (14) species were found at all microenvironments.