IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Hanseniaspora in wine-making: their genetic modification and potential role in acid modulation

Hanseniaspora in wine-making: their genetic modification and potential role in acid modulation

Abstract

Hanseniaspora spp. are one of the most common yeast isolates in vineyards and wineries and play an important role in wine-making.  We explored the impact of an apiculate yeast Hanseniaspora occidentalis strain as a co-partner with Saccharomyces cerevisiae in a sequential-type mixed-culture fermentation of Muscaris grape must.  Like with other fermentation trials using Hanseniaspora strains, a significant increase in ethyl acetate was observed, but most intriguing, was the almost complete abolition of malic acid in the wine.  Compared to the pure S. cerevisiae inoculum there was also a marked increase in the concentrations of the other acetate esters. Modulation of some of the varietal elements like rose oxide was also observed.  Identifying and confirming the genes involved in malic acid utilization and aroma formation would require the development of gene modification tools: a feature not existing yet in H. occidentalis. We have recently developed gene-modifying tools in Hanseniaspora uvarum where we knocked-out the two alleles of the alcohol acetyltransferase gene (HuATF) resulting in a yeast with much lower acetate ester capabilities. This was the first successful attempt to genetically remove a gene from Hanseniaspora and paves the way for further gene-to-function studies in this apiculate yeast
genus

DOI:

Publication date: June 23, 2022

Issue: IVAS 2022

Type: Article

Authors

Van Wyk Niel¹, Badura Jennifer¹, Scansani Stefano¹,³, Pretorius Isak S.², Rauhut Doris1, Von Wallbrunn Christian¹

¹Department of Microbiology and Biochemistry, Hochschule Geisenheim University, Geisenheim, Germany
²ARC Centre of Excellence in Synthetic Biology, Department of Molecular Sciences, Macquarie University, Sydney, Australia
³Institute of Applied Microbiology, Justus-Liebig-University, Giessen, Germany

Contact the author

Keywords

Hanseniaspora, GMO, malic acid

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

Diagnosis of soil quality and evaluation of the impact of viticultural practices on soil biodiversity in a vineyard in southwestern France

Viticulture is facing two major changes – climate change and agroecological transition. In both cases, soil quality is seen as a lever to move towards a more sustainable viticulture. However, soil biological quality is little considered in the implementation of viticultural practices. Gascogn’Innov (2017-2022) is an Operational Group funded by the European Innovation Partnership for Agriculture. As such, it brings together winegrowers from the south-west of France, scientists, advisors and technicians, around a project focused on viticultural soil biological functioning and the design of technical routes more respectful toward soil heritage. To achieve this, the project aims to acquire references on the impact of viticultural practices on soil biology from a dynamic way, and to test a methodology to integrate information provided by the soil bioindicators to manage farming systems. A set of indicators of soil biological quality are evaluated in the project: microorganisms (bacteria and fungi abundance and diversity), fauna (abundance and diversity of nematodes and earthworms), physico-chemical characteristics, soil structure assessment and degradation rate of organic matter. Based on a network of 13 plots that have been subject to an initial diagnosis in 2017, several agronomical practices to restore soil fertility are experimented to redesign the cropping system (for instance plant cover, organic matter inputs, reduction of herbicides, mineral fertilizers). System redesign was made in collaboration by winegrowers and an interdisciplinary group of experts (agronomists, biologists). Several indicators are measured on vine and soil at each vintage to assess vine health and productivity. At the end of the project (2021), a final diagnosis was carried out. Gascogn’Innov allowed to create a regional database on the quality of wine-growing soils, which permitted to evaluate the effect of practices according to soil types. Especially, decreasing the intensity of tillage and increasing the duration and diversity of grass coverage tends to increase the abundance of all the organisms studied. This project confirmed the value of soil biological quality indicators to drive the sustainability of practices, but also highlighted the key-role of expertise, in both agronomy and soil biology, to help winegrowers understand and appropriate their soil quality diagnoses.

VINIoT – Precision viticulture service

The project VINIoT pursues the creation of a new technological vineyard monitoring service, which will allow companies in the wine sector in the SUDOE space to monitor plantations in real time and remotely at various levels of precision. The system is based on spectral images and an IoT architecture that allows assessing parameters of interest viticulture and the collection of data at a precise scale (level of grape, plant, plot or vineyard) will be designed. In France, three subjects were specifically developed: evaluation of maturity, of water stress, and detection of flavescence dorée. For the evaluation of maturity, it has been decided first to work at the berry scale in the laboratory, then at the bunch scale and finally in the vineyard. The acquisition of the spectral hyperstal image as well as the reference analyzes to measure the maturity, were carried out in the laboratory after harvesting the berries in a maturity monitoring context. This work focuses on a case study to predict sugar content of three different grape varieties: Syrah, Fer Servadou and Mauzac. A robust method called Roboost-PLSR, developed in the framework of this work (Courand et al., 2022), to improve prediction model performance was applied on spectra after the acquirement of hyperspectral images. Regarding the evaluation of water stress, to work with a significant variability in terms of water status, it has been worked first with potted plants under 2 different water regimes. The facilities have allowed the supervision of irrigation and micro-climatic conditions. The regression models on agronomic variables (stomatal conductance, water potential, …) are studied. To detect flavescence dorée, the experimental plan has consisted of work at leaf scale in the laboratory first, and then in the field. To detect the disease from hyper-spectral imaging, a combination of multivariate curve resolution-alternating least squares (MCR-ALS) and factorial discriminant analysis (FDA) was proposed. This strategy proved the potential towards the discrimination of healthy and infected leaves by flavescence dorée based on the use of hyperspectral images (Mas Garcia et al., 2021).

Metabolomic discrimination of grapevine water status for Chardonnay and Pinot noir

Water status impact in viticulture has been widely explored, as it strongly affects grapevine physiology and grape chemical composition. It is considered as a key component of vitivinicultural terroir. Most of the studies concerning grapevine water status have focused on either physiological traits, or berry compounds, or traits involved in wine quality. Here, the response of grapevine to water availability during the ripening period is assessed through non-targeted metabolomics analysis of grape berries by ultra-high resolution mass spectrometry. The grapevine water status has been assessed during 2 consecutive years (2019 & 2020), through carbon isotope discrimination on juices from berries collected at maturity (21.5 brix approx.) for 2 Vitis vinifera cv. Pinot noir (PN) and Chardonnay (CH). A total of 220 grape juices were collected from 5 countries worldwide (Italy; Argentina; France; Germany; Portugal). Measured δ13C (‰) varied from -28.73 to -22.6 for PN, and from -28.79 to -21.67 for CH. These results also clearly revealed higher water stress for the 2020 vintage. The same grape juices have been analysed by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) and Liquid Chromatography coupled to Mass Spectrometry (LC-qTOF-MS), leading to the detection of up to 4500 CHONS containing elemental compositions, and thus likely tens of thousands of individual compounds, which include fatty acids, organic acids, peptides, phenolics, also with high levels of glycosylation. Multivariate statistical analysis revealed that up to 160 elemental compositions, covering the whole range of detected masses (100 –1000 m/z), were significantly correlated to the observed gradients of water status. Examples of chemical markers, which are representative of these complex fingerprints, include various derivatives of the known abscisic acid (ABA), such as phaesic acid or abscisic acid glucose ester, which are significantly correlated with higher water stress, regardless of the variety. Cultivar-specific behaviours could also be identified from these fingerprints. Our results provide an unprecedented representation of the metabolic diversity, which is involved in the water status regulation at the grape level, and which could contribute to a better knowledge of the grapevine mitigation strategy in a climate change context.

Genotypic variability in root architectural traits and putative implications for water uptake in grafted grapevine

Root system architecture (RSA) is important for soil exploration and edaphic resources acquisition by the plant, and thus contributes largely to its productivity and adaptation to environmental stresses, particularly soil water deficit. In grafted grapevine, while the degree of drought tolerance induced by the rootstock has been well documented in the vineyard, information about the underlying physiological processes, particularly at the root level, is scarce, due to the inherent difficulties in observing large root systems in situ. The objectives of this study were to determine genetic differences in the root architectural traits and their relationships to water uptake in two Vitis rootstocks genotypes (RGM, 140Ru) differing in their adaptation to drought. Young rootstocks grafted upon the Riesling variety were transplanted into cylindrical tubes and in 2D rhizotrons under two conditions, well watered and moderate water stress. Root traits were analyzed by digital imaging and the amount of transpired water was measured gravimetrically twice a week. Root phenotyping after 30 days reveal substantial variation in RSA traits between genotypes despite similar total root mass; the drought-tolerant 140Ru showed higher root length density in the deep layer, while the drought-sensitive RGM was characterised by shallow-angled root system development with more basal roots and a larger proportion of fine roots in the upper half of the tube. Water deficit affected canopy size and shoot mass to a greater extent than root development and architectural-related traits for both 140Ru and RGM, suggesting vertical distribution of roots was controlled by genotype rather than plasticity to soil water regime. The deeper root system of 140Ru as compared to RGM correlated with greater daily water uptake and sustained stomata opening under water-limited conditions but had little effect on above-ground growth. Our results highlight that grapevine rootstocks have constitutively distinct RSA phenotypes and that, in the context of climate change, those that develop an extensive root network at depth may provide a desirable advantage to the plant in coping with reduced water resources.

Evaluation of climate change impacts at the Portuguese Dão terroir over the last decades: observed effects on bioclimatic indices and grapevine phenology

In the last decades the growers of the Portuguese Dão winegrowing region (center of Portugal) are experiencing changes in climate that are influencing either grape phenology berry health and ripening. Aiming to study the relationships between climate indices (CI), seasonal weather and grapevine phenology, in this work long-term climate and phenological data collected at the experimental vineyard of the Portuguese Dão research centre between 1958 and 2019 (61 years) for the red variety Touriga Nacional, was analyzed. The trends over time for the classical temperature-based indices (Growing Season Temperature – GST -, Growing Degree Days – GDD, Huglin Index – HI and Cool Night Index – CI) presented a significantly positive slope while the Dryness Index (DI) showed a negative trend over the last 61 years. Regarding grapevine phenology, an average advance of 4.5 days per decade in the harvest day was observed throughout the last 61 years. Consequently, the weather conditions during the ripening period have changed, showing an increasing trend over time in the average temperature (higher magnitude in the maximum than in the minimum temperature) and a decrease in the accumulated rainfall. A regression analysis showed that ~50% of harvest date variability over years was explained by the temperature-based indices variability. These observed effects of climate change on bioclimatic indices and corresponding anticipation of harvest date can still be considered advantageous for the Dão terroir as it allows to achieve an optimal berry ripening before the common equinox rains and, therefore, avoid the potential negative impacts of the rainfall on berry health and composition.