Macrowine 2021
IVES 9 IVES Conference Series 9 Improvement of non-Saccharomyces yeast dominance during must fermentation by using spontaneous mutants resistant to SO2, EtOH and high pressure of CO2

Improvement of non-Saccharomyces yeast dominance during must fermentation by using spontaneous mutants resistant to SO2, EtOH and high pressure of CO2

Abstract

AIM: A genetic study of four wine T. delbrueckii strains was done. Spore clones free of possible recessive growth‐retarding alleles with enhanced resistance to winemaking stressing conditions were obtained from these yeasts.

METHODS: The genetic marker of resistance to cycloheximide (cyhR) allows easy monitoring of the new mutants obtained from these yeasts. Identity of the high pressure resistant (HPR) mutants was verified by analysis of cell morphology, killer phenotype, cyhR marker, presence of viral dsRNA, RFLPs of mtDNA, and sequencing of Internal Transcribed Spacer ofribosomal DNA (ITS).

RESULTS: T. Delbrueckii mutants were isolated from some spore clones. Papillae resistant to SO2 were isolated. Subsequently, new spontaneous mutants capable of growing on YEPD plates with 10% ethanol were isolated. Rosé sparkling wine (cava) was made using these mutants. Two mutants, with the best fermentation kinetics and closest to the reference yeast Sc 85R4, were isolated from some bottles with high CO2 pressure and some were selected there after (Td MutHP41 and Td MutHP42). They had better fermentation kinetics and dominance than their parental yeast. Td MutHP41 showed great improvement for industrial base wine fermentation with respect to its parental yeast. Re-isolation and selection procedure to obtain new reinforced HPR mutants from previously selected HPR mutants was not a sound strategy to continue improving the fermentative capability of T. delbrueckii under high CO2 pressure. Continuous shaking during inocula preparation further improved the fermentative capability of T. delbrueckii yeasts.

CONCLUSIONS: Isolation of spontaneous mutants resistant to SO2 and ethanol seems to be a good strategy to slightly improve the fermentative efficiency of T. delbrueckii in must and base wine. The new mutants were genetically stable enough to be considered for industrial production, and their fermentative capability was further improved by continuously supplying oxygen during the conditioning stage before yeast culture inoculation in base wine.

DOI:

Publication date: September 10, 2021

Issue: Macrowine 2021

Type: Article

Authors

Alberto Martínez Brígido 1, Rocío Velázquez 1, Joaquín Bautista-Gallego 1, Emiliano Zamora 2, Manuel Ramírez 1

1 Departamento de Ciencias Biomédicas, Universidad de Extremadura, 06006 Badajoz, Spain.
2 Estación Enológica, Junta de Extremadura. 

Contact the author

Keywords

Torulaspora delbrueckii; wine fermentation, sporeclone; sparkling wine; ethanol resistance; SO2 resistance; pressure resistance

Citation

Related articles…

Effects of graft quality on growth and grapevine-water relations

Climate change is challenging viticulture worldwide compromising its sustainability due to warmer temperatures and the increased frequency of extreme events. Grafting Vitis vinifera L.

Photoselective shade films affect grapevine berry secondary metabolism and wine composition

Grapevine physiology and production are challenged by forecasted increases in temperature and water deficits. Within this scenario, photoselective overhead shade films are promising tools in warm viticulture areas to overcome climate change related factors. The aim of this study was to evaluate the vulnerability of ‘Cabernet Sauvignon’ grape berry to solar radiation overexposure and optimize shade film use for berry integrity. A randomized complete block design field study was conducted across two years (2020-2021) in Oakville, Napa Valley, CA, with four shade films (D1, D3, D4, D5) differing in the percent of radiation spectra transmitted and compared to an uncovered control (C0). Integrals for gas exchange parameters and mid-day stem water potential were unaffected by the shade films in 2020 and 2021. By harvest, berries from uncovered and shaded vines did not differ in their size or primary metabolism in either year. Despite precipitation exclusion during the dormant season in the shaded treatments, yield did not differ between them and the control in either season. In 2020, total skin anthocyanins (mg/g fresh mass) in the shaded treatments was greater than C0 during berry ripening and at harvest. Conversely, flavonol concentrations in 2020 were reduced in shaded vines compared to C0. The 2020 growing season highlighted the impact of heat degradation on flavonoids. Flavonoid concentrations in 2021 increased until harvest while flavonoid degradation was apparent from veraison to harvest in 2020 across shaded and control vines. Wine analyses highlighted the importance of light spectra to modify wine composition. Wine color intensity, tonality and anthocyanin values were enhanced in D4 whereas antioxidant properties were enhanced in C0 and D5 wines. Altogether, our results highlighted the need of new approaches in warm viticulture areas given the impact that composition of light has on berry and wine quality.

Adaptation to soil and climate through the choice of plant material

Choosing the rootstock, the scion variety and the training system best suited to the local soil and climate are the key elements for an economically sustainable production of wine. The choice of the rootstock/scion variety best adapted to the characteristics of the soil is essential but, by changing climatic conditions, ongoing climate change disrupts the fine-tuned local equilibrium. Higher temperatures induce shifts in developmental stages, with on the one hand increasing fears of spring frost damages and, on the other hand, ripening during the warmest periods in summer. Expected higher water demand and longer and more frequent drought events are also major concerns. The genetic control of the phenotypes, by genomic information but also by the epigenetic control of gene expression, offers a lot of opportunities for adapting the plant material to the future. For complex traits, genomic selection is also a promising method for predicting phenotypes. However, ecophysiological modelling is necessary to better anticipate the phenotypes in unexplored climatic conditions Genetic approaches applied on parameters of ecophysiological models rather than raw observed data are more than ever the basis for finding, or building, the ideal varieties of the future.

Effect of regulated deficit irrigation regime on amino acids content of Monastrell (Vitis vinifera L.) grapes

Irrigation is an important practice to influence vine quality, especially in Mediterranean regions, characterized by hot summers and severe droughts during the growing season. This study focused on deficit irrigation regime influence on amino acids composition of Monastrell grapevines under semiarid conditions (Albacete, Southeastern of Spain). In 2019, two treatments were applied: non-irrigation (NI) and regulated deficit irrigation (RDI), watered at 30% of the estimated crop evapotranspiration from fruit set to onset of veraison. Grape amino acids content was analyzed by HPLC. Berries from non-irrigated vines showed higher concentration of several amino acids, such as tryptophan (73%), arginine (70%), lysine (36%), isoleucine (27%), and leucine (21%), compared to RDI grapes. Arginine is, together with ammonium ion, the principal nitrogen source for yeasts during the alcoholic fermentation; while isoleucine, tryptophan, and leucine are precursors of fermentative volatile compounds, key compounds for wine quality. Moreover, NI treatment increased in a 14% the total amino acids content in grapes compared to RDI treatment. The reported effects might be because yield was 70% higher in RDI vines than in the NI ones and, therefore, the sink demand was increased in the irrigated vines. In addition, NI vines suffered more severe water stress and it is known that the amino acids synthesis and accumulation can be influenced by the plant response to stress. According to the results, the irrigation regime showed effect on amino acids concentration in Monastrell grapes under semiarid conditions. Grapes from non-irrigated vines showed a higher content of several amino acids relevant to the fermentative process and to the wine aroma compounds formation. It is demonstrated that the final content of nitrogen-related components in grapes is influenced by the irrigation regime. The convenience of the irrigation strategy to suggest will depend on the desired wine style and the target yield levels.

Effects of organic mulches on the soil environment and yield of grapevine

Farming management practices aiming at conserving soil moisture have been developed in arid and semiarid-areas facing water scarcity problems. Organic mulching is an effective method to manipulate the crop-growing microclimate increasing crop yield by controlling soil temperature, and retaining soil moisture by reducing soil evaporation. In this sense, the effectiveness of different organic mulching materials (straw mulch and grapevine pruning debris) applied within the row of a vineyard was evaluated on the soil and on the vine in a Tempranillo vineyard located in La Rioja (Spain). Organic mulches were compared with a traditional bare soil management technique (based on the use of herbicides to avoid weed incidence). Mulching coverages favourably influenced the soil water retention throughout all the grapevine vegetative cycle. However, the soil-moisture variation was not the same under different mulching materials, being the straw mulch (SM) the one that retained more water in comparison with grapevine pruning debris (GPD) based-cover. The changes of soil moisture in the upper surface layer (0–10 cm) were highly dynamic, probably due to water vapour fluxes across the soil-atmospheric interface. However, both, SM and GPD reduced these fluctuations as compared with bare soils. A similar trend occurred with soil temperature. Both organic mulches altered soil temperature in comparison with bare soil by reducing soil temperature in summer and raising it in winter. Moreover, the same buffering effect for the temperature on the covered soil also remains in the deeper layers. To conclude, we could see that organic mulching had a positive impact on soil-moisture storage and soil temperature and the extent of this effect depends on the type of mulching materials. These changes led to higher rates of photosynthesis and stomatal conductivity compared to bare soils, also favouring crop growth and grape yields.