Biocontrol of Alternaria alternata in cold-stored table grapes using psychrotrophic yeasts and bioactive compounds of natural sources.

Alternaria alternata is a common fungal pathogen causing postharvest decay in table grapes. This study addressed the potential of autochthonous yeasts and bioactive compounds of natural sources to act as biocontrol agents (BCAs) against A. alternata in cold-stored table grapes. With this purpose, 19 yeast capable of growing at 0-1 °C were isolated from the surface of Red Globe table grapes. These isolates, along with the pre-isolated strain Metschnikowia pulcherrima RCM2, were evaluated as BCAs in wounded berries. From these results, six yeast isolates were pre-selected to be combined with bioactive compounds of natural sources, like phenolic compounds (PCs) of side streams of wine industry, including bunch stem extract (BSE) (5-25 %), and cane extract (CE) (5-25 %), and functional polysaccharides from shrimp waste such as chitosan (CH) (0.5 %). Then, the biocontrol efficacy of combined treatments beyond individual ones was compared. The results revealed that 4 yeast isolates, namely M. pulcherrima RCM2 and ULA146, and Aureobasidium pullulans FUL14 and FUL18, were the most effective. However, when combined with the natural bioactive compounds, their efficacy against A. alternata did not increase significantly. Notably, ULA146 and FUL18 demonstrated a biocontrol efficacy of 36-37 %, comparable to that of the treatment with commercial doses of SO2, which only showed a 27 % reduction in the lesion diameter. These findings highlight the potential of using psychrotrophic yeasts as BCAs against A. alternata in cold-stored table grapes. Combining these yeast strains with BSE, CE and CH did not increase BCAs efficacy against this pathogen at the concentrations tested. The development of effective biocontrol strategies for A. alternata could contribute to reducing reliance on chemically synthesized fungicides, promoting sustainable practices, aiming to improve the quality and safety of cold-stored table grapes.

Evaluation of different nitrogen sources on growth and fermentation performance for enhancing ethanol production by wine yeasts.

The utilization of grape juice from low oenological value grape varieties for bioethanol production represent an alternative for diversification and value addition in viticulture. Optimizing Very High Gravity (VHG) fermentation can significantly increase ethanol productivity while reducing water and energy consumption. In this study, the impact of different nitrogen sources on growth and fermentative performance of locally selected yeast strains was investigated. Five yeast strains of species Saccharomyces cerevisiae and Zygosaccharomyces rouxii were cultured in both synthetic culture media and natural grape juice supplemented with ammonium sulfate (NH), yeast extract (YE), Fermaid K (FERM), and urea (U) at varying concentrations. Due to the very low fermentation rate, the Z. rouxii strain was excluded from the selection. The results obtained in synthetic medium showed that nitrogen sources that promoted growth (NH and YE) had minimal effects on fermentative performance and were highly dependent on the specific yeast strain. However, the combination of urea and ammonium favored the rate of sugar consumption. When validated in natural grape juice, urea combined with ammonium (U + NH 300 + 75 mg/L) improved both growth parameters and ethanol yield. Doubling the concentration (U + NH 600 + 150 mg/L) further enhanced sugar consumption and ethanol production while reducing unwanted by-products. The combined use of urea and ammonium exhibited a synergistic effect, making it a cost-effective nitrogen supplement for VHG fermentations.

Generation of a Non-Transgenic Genetically Improved Yeast Strain for Wine Production from Nitrogen-Deficient Musts.

The yeast Saccharomyces cerevisiae is the main species responsible for the process that involves the transformation of grape must into wine, with the initial nitrogen in the grape must being vital for it. One of the main problems in the wine industry is the deficiency of nitrogen sources in the grape must, leading to stuck or sluggish fermentations, and generating economic losses. In this scenario, an alternative is the isolation or generation of yeast strains with low nitrogen requirements for fermentation. In the present study, we carry out a genetic improvement program using as a base population a group of 70 strains isolated from winemaking environments mainly in Chile and Argentina (F0), making from it a first and second filial generation (F1 and F2, respectively) based in different families and hybrids. It was found that the trait under study has a high heritability, obtaining in the F2 population strains that consume a minor proportion of the nitrogen sources present in the must. Among these improved strains, strain "686" specially showed a marked drop in the nitrogen consumption, without losing fermentative performance, in synthetic grape must at laboratory level. When using this improved strain to produce wine from a natural grape must (supplemented and non-supplemented with ammonium) at pilot scale under wine cellar conditions, a similar fermentative capacity was obtained between this strain and a widely used commercial strain (EC1118). However, when fermented in a non-supplemented must, improved strain "686" showed the presence of a marked floral aroma absent for EC1118 strain, this difference being probably a direct consequence of its different pattern in amino acid consumption. The combination of the capacity of improved strain "686" to ferment without nitrogen addition and produce floral aromas may be of commercial interest for the wine industry.

Biodiversity of Saccharomyces cerevisiae populations in Malbec vineyards from the "Zona Alta del Río Mendoza" region in Argentina.

The "Zona Alta del Río Mendoza" (ZARM) is the major Malbec grape viticulture region of Argentina. The aim of the present study was to explore Saccharomyces cerevisiae biodiversity in ZARM vineyards. Interdelta PCR and RFLP mtDNA molecular markers were applied to differentiate S. cerevisiae strains. The presence of commercial strains on ZARM vineyards was also assessed. Our results reveal a highly diverse, but genetically closely related, S. cerevisiae population (containing more than 190 molecular patterns among 590 S. cerevisiae isolates). According to the S. cerevisiae strain diversity found in vineyards, they were classified as vineyards with high and low polymorphic S. cerevisiae populations. Six vineyards showed a high polymorphic population, with more than 20 different S. cerevisiae molecular patterns. S. cerevisiae populations in these vineyards were diverse and irregularly distributed, with different strains in each vineyard site. Low polymorphic S. cerevisiae population vineyards displayed very low yeast diversity, with only 9 to 10 different S. cerevisiae strains and presence of two commercial strains widely distributed. Population diversity estimators were calculated to determine the population structure of S. cerevisiae in the ZARM vineyards. The obtained values support the hypothesis that the eight sampled subpopulations come indeed from a larger population.