Macrowine 2021
IVES 9 IVES Conference Series 9 Screening of soil yeasts with fermentative capacity from the antarctic continent for their application in the wine industry

Screening of soil yeasts with fermentative capacity from the antarctic continent for their application in the wine industry

Abstract

AIM: In the last years, many wineries are increasing experimentation to produce more distinguishable beverages. In this sense, the reduction of the fermentation temperature could be a useful tool because it preserves volatile compounds and prevents wines from browning, particularly in the case of white wines. However, low fermentation temperatures usually produce an early stop in the fermentation. Hence, the employment of new yeast strains able to operate at low temperatures could solve this problem, giving rise to different characteristics in wines. The Antarctic Continent is a crucial source for the isolation of new microorganisms and bioactive metabolites, given the competitive conditions of this environment with low temperatures, scarce carbon sources and high UV radiation. Considering this, the objective of this study was the isolation and characterization of fermentative yeast from the Antarctic Continent with potential for use in the wine industry.

METHODOLOGY: Six soil samples collected in Fildes Bay, west of King George Island and three soil samples from King George Island were processed for yeast isolation. Samples were suspended in sterile water and dilutions of each suspension were inoculated onto yeast medium (YM) agar plates with antibiotics, which were then incubated at 4, 10, and 18 °C until visible colony growth. Colonies with a non-filamentous appearance were selected, which were reseeded on YM agar without antibiotics. Alcohol tolerance was performed using concentrations of 3, 6, or 9% alcohol. Later on, sugar tolerance was analyzed using fructose and glucose in a 1:1 proportion; with 5, 10, 15, 20, or 25% of sugar in the medium. Those isolates with microscopic characteristics of interest were selected to determine fermentative activity in vitro using a simple colorimetric assay with phenol red, as a pH indicator. To differentiate the isolates, and discard replicates, a fingerprinting assay with arbitrary primers was performed. Identification of the isolates was carried out using PCR and ITS region primers with BLAST bioinformatics tools.

RESULTS: Nine soil samples collected from the Antarctica were processed for yeast isolation. We obtained 125 yeasts from the soil samples, with a growth temperature of 10ºC. Overall, 25 yeasts have fermentative activity and are able to tolerate a culture medium with at least 20% glucose and up to 6% of ethanol. The isolates were also characterized by optical microscopy and fingerprinting using PCR with arbitrary primers to discard identical strains and allowed us to discriminate 10 unique strains with fermentative capacity from the 25 isolates. To determine the identity of the isolated yeasts, the amplification and sequence of the 18S RNA was performed.

CONCLUSION:

The Antarctic continent has proven to be a source of fermentative yeasts with high potential for their use in the wine industry.

DOI:

Publication date: September 3, 2021

Issue: Macrowine 2021

Type: Article

Authors

Laura Navarro, Cristina Úbeda, Mariona Gil i Cortiella, Ana Gutierrez, Gino Corsini, Nancy Calisto

Biomedical Sciences Institute, Health Sciences Faculty, Autonomous University of Chile, Santiago, Chile., Nutrition and Food Sciences Department, Faculty of Pharmacy, University of Seville, Seville, Spain. Applied Chemical Sciences Institute, Autonomous University of Chile, Santiago, Chile. Biomedical Sciences Institute, Health Sciences Faculty, Autonomous University of Chile, Santiago, Chile. Biomedical Sciences Institute, Health Sciences Faculty, Autonomous University of Chile, Santiago, Chile. Chemical engineering department, Faculty of Engineering, Magallanes University, Punta Arenas, Chile. Biomedical Sciences Institute, Health Sciences Faculty, Autonomous University of Chile, Santiago, Chile.

Contact the author

Keywords

antarctic yeast, low temperature fermentation, yeast isolation, yeast characterization

Citation

Related articles…

Characterization of variety-specific changes in bulk stomatal conductance in response to changes in atmospheric demand and drought stress

In wine growing regions around the world, climate change has the potential to affect vine transpiration and overall vineyard water use due to related changes in atmospheric demand and soil water deficits. Grapevines control their transpiration in response to a changing environment by regulating conductance of water through the soil-plant-atmosphere continuum. Most vineyard water use models currently estimate vine transpiration by applying generic crop coefficients to estimates of reference evapotranspiration, but this does not account for changes in vine conductance associated with water stress, nor differences thought to exist between varieties. The response of bulk stomatal conductance to daily weather variability and seasonal drought stress was studied on Cabernet-Sauvignon, Merlot, Tempranillo, Ugni blanc, and Semillon vines in a non-irrigated vineyard in Bordeaux France. Whole vine sap flow, temperature and humidity in the vine canopy, and net radiation absorbed by the vine canopy were measured on 15-minute intervals from early July through mid-September 2020, together with periodic measurement of leaf area, canopy porosity, and predawn leaf water potential. From this data, bulk stomatal conductance was calculated on 15-minute intervals, and multiple regression analysis was performed to identify key variables and their relative effect on conductance. Attention was focused on addressing multicollinearity and time-dependency in the explanatory variables and developing regression models that were readily interpretable. Variability of vapor pressure deficit over the day, and predawn water potential over the season explained much of the variability in conductance, with relative differences in response coefficients observed across the five varieties. By characterizing this conductance response, the dynamics of vine transpiration can be better parameterized in vineyard water use modeling of current and future climate scenarios.

Effect of partial net shading on the temperature and radiation in the grapevine canopy, consequences on the grape quality of cv. Gros Manseng in PDO Pacherenc-du-vic-Bilh

As elsewhere, southwestern France vineyards face more recurrent summer heat waves these last years. Among the possibilities of adaptation to this climate changing parameter, the use of net shading is a technique that allow for limiting canopy exposure to radiations. In this trial, we tested net shading installed on one face of the canopy, on a north-south row-oriented plot of cv. Gros Manseng trained on VSP system in the PDO Pacherenc-du-Vic-Bilh. The purpose was to characterize the effects on the ambient canopy temperatures and radiations during the season and to observe the consequences on the composition of grapes and wines. Two sorts of net were used with two levels of obstruction (50% and 75%) of the photosynthesis active radiation (PAR). They have been installed on the west side of the canopy and compared to a netless control. Temperature and PAR sensors registered hourly data during the season. On specific summer day (hot and sunny) manual measurements took also place on bunches (temperature) and in different spots of the canopy (PAR). The results showed that, on clear days, the radiation is lowered by the shade nets respecting the supplier criteria. The effects on the ambient canopy temperature were inconstant on this plot when we observed the data from the global period of shading between fruit set and harvest. However, during hot days (>30°C), the temperature in the canopy was reduced during afternoon and the temperature of the bunch surface was reduced as well comparing to the control. A decrease of the maturity parameters of the berries, sugar and acidity, was also observed. Concerning the wine aromatic potential, no differences clearly appeared.

Amino nitrogen content in grapes: the impact of crop limitation

As an essential element for grapevine development and yield, nitrogen is also involved in the winemaking process and largely affects wine composition. Grape must amino nitrogen deficiency affects the alcoholic fermentation kinetics and alters the development of wine aroma precursors. It is therefore essential to control and optimize nitrogen use efficiency by the plant to guarantee suitable grape nitrogen composition at harvest. Understanding the impact of environmental conditions and cultural practices on the plant nitrogen metabolism would allow us to better orientate our technical choices with the objective of quality and sustainability (less inputs, higher efficiency). This trial focuses on the impact of crop limitation – that is a common practice in European viticulture – on nitrogen distribution in the plant and particularly on grape nitrogen composition. A wide gradient of crop load was set up in a homogeneous plot of Chasselas (Vitis vinifera) in the experimental vineyard of Agroscope, Switzerland. Dry weight and nitrogen dynamics were monitored in the roots, trunk, canopy and grapes, during two consecutive years, using a 15N-labeling method. Grape amino nitrogen content was assessed in both years, at veraison and at harvest. The close relationship between fruits and roots in the maintenance of plant nitrogen balance was highlighted. Interestingly, grape nitrogen concentration remained unchanged regardless of crop load to the detriment of the growth and nitrogen content of the roots. Meanwhile, the size and the nitrogen concentration of the canopy were not affected. Leaf gas exchange rates were reduced in response to lower yield conditions, reducing carbon and nitrogen assimilation and increasing intrinsic water use efficiency. The must amino nitrogen profiles could be discriminated as a function of crop load. These findings demonstrate the impact of plant balance on grape nitrogen composition and contribute to the improvement of predictive models and sustainable cultural practices in perennial crops.

De novo Vitis champinii whole genome assembly allows rootstock-specific identification of potential candidate genes for drought and salt tolerance

Vitis champinii cultivars Ramsey and Dog-ridge are main choices for rootstocks to adapt viticulture in semi-arid and arid regions thanks to their distinctive tolerance to drought and salinity. However, genetic studies on non-vinifera rootstocks have heavily relied on the grapevine (Vitis vinifera) reference genome, which difficulted the assessment of the genetic variation between rootstock species and grapevines. In the present study, this limitation is addressed by introducing a novo phased genome assembly and annotation of Vitis champinii. This new Vitis champinii genome was employed as reference for mapping RNA-seq reads from the same species under drought and salt stresses, and for comparison the same reads were also mapped to the Vitis vinifera PN40024.V4 reference genome. A significant increase in alignment rate was gained when mapping Vitis champinii RNA-seq reads to its own genome, compared to the Vitis vinifera PN40024.V4 reference genome, thus revealing the expression levels of genes specific to Vitis champinii. Moreover, differences in coding sequences were observed in ortholog genes between Vitis champinii and Vitis vinifera, which therefore challenges previous differential expression analyses performed between contrasting Vitis genotypes on the same gene from the Vitis vinifera genome. Genes with possible implications in drought and salt tolerance have been identified across the genome of Vitis champinii, and the same genomic data can potentially guide the discovery of candidate genes specific from Vitis champinii for other traits of interest, therefore becoming a valuable resource for rootstock breeding designs, specially towards increased drought and salinity due to climate change.

Sustaining wine identity through intra-varietal diversification

With contemporary climate change, cultivated Vitis vinifera L. is at risk as climate is a critical component in defining ecologically fitted plant materiel. While winegrowers can draw on the rich diversity among grapevine varieties to limit expected impacts (Morales-Castilla et al., 2020), replacing a signature variety that has created a sense of local distinctiveness may lead to several challenges. In order to sustain wine identity in uncertain climate outcomes, the study of intra-varietal diversity is important to reflect the adaptive and evolutionary potential of current cultivated varieties. The aim of this ongoing study is to understand to what extent can intra-varietal diversity be a climate change adaptation solution. With a focus on early (Sauvignon blanc, Riesling, Grolleau, Pinot noir) to moderate late (Chenin, Petit Verdot, Cabernet franc) ripening varieties, data was collected for flowering and veraison for the various studied accessions (from conservatory plots) and clones. For these phenological growing stages, heat requirements were established using nearby weather stations (adapted from the GFV model, Parker et al., 2013) and model performances were verified. Climate change projections were then integrated to predict the future behaviour of the intra-varietal diversity. Study findings highlight the strong phenotypic diversity of studied varieties and the importance of diversification to enhance climate change resilience. While model performances may require improvements, this study is the first step towards quantifying heat requirements of different clones and how they can provide adaptation solutions for winegrowers to sustain local wine identity in a global changing climate. As genetic diversity is an ongoing process through point mutations and epigenetic adaptations, perspective work is to explore clonal data from a wide variety of geographic locations.