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IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Optimization and validation of a fully automated HS-SPME method for determination of VCCs and its application in wines submitted to accelerated ageing

Optimization and validation of a fully automated HS-SPME method for determination of VCCs and its application in wines submitted to accelerated ageing

Abstract

Wine aroma is a complex gaseous mixture composed of various compounds; some of these molecules derive directly from the grapes while most of them are released and synthetized during fermentation or are due to ageing reactions. Among the latter class of compounds, carbonyls are the principal products of oxidation reactions which take place during the storing time. Volatile carbonyl compounds (VCCs) are related to aromatic nuances of vanilla, caramel, butter, honey, potato, orange, lemon, violets, cider and plum, which are pleasant scents characteristics of oxidized wines. However, apart from cases where it is a deliberate process, oxidation is commonly undesired and the presence of a relevant content of carbonyls is related to aroma defects. Because of that, monitoring the concentration of VCCs could be added as a quality control for the evaluation of a complete fermentation, correct winemaking style, and proper bottling and storage. In this research an HS-SPME method1 was optimized and validated with the aim to be used as a tool to achieve this goal. The use of a solvent-free extraction allowed to maximize the coherence to the Green Analytical Chemistry principles with a simultaneous achievement in performance, reliability and robustness. In this method, all sample preparation steps were automated using the autosampler minimizing the human time consumption to enhance the scalability to routine analysis. As many as 46 VCCs (mainly linear aldehydes, Strecker aldehydes, unsaturated aldehydes, ketones, and many other) were the analytes under investigation. All compounds showed a good linearity spanning from approximately 0.1 to 100 µg/L (R2>0.99). Intra-day and 5 days inter-days repeatability showed an RSD

DOI:

Publication date: June 23, 2022

Issue: IVAS 2022

Type: Article

Authors

Piergiovanni Maurizio1, Carlin Silvia2, Lotti Cesare2, Vhrovsek Urska2 and Mattivi Fulvio1,2

1Center Agriculture Food Environment (C3A), University of Trento, via Edmund Mach 1, San Michele all’Adige (TN) Italy
2Center Research and Innovation, Edmund Mach Foundation, Italy3Affiliation of the third 

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Keywords

Carbonyls, oxidation, accelerated ageing, HS-SPME, quality control

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IVAS 2022 | IVES Conference Series

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Deconstructing the soil component of terroir: from controversy to consensus

Wine terroir describes the collectively recognized relation between a geographical area and the distinctive organoleptic characteristics of the wines produced in it. The overriding objective in terroir studies is therefore to provide scientific proof relating the properties of terroir components to wine quality and typicity. In scientific circles, the role of climate (macro-, meso- and micro-) on grape and wine characteristics is well documented and accepted as the most critical. Moreover, there has been increasing interest in recent years about new elements with possible importance in shaping wine terroir like berry/leaf/soil microbiology or even aromatic plants in proximity to the vineyard conferring flavors to the grapes. However, the actual effect of these factors is also dependent on complex interactions with plant material (variety/clone, rootstock, vine age) and with human factors.
The contribution of soil, although a fundamental component of terroir and extremely popular among wine enthusiasts, remains a much-debated issue among researchers. The role of geology is probably the one mostly associated by consumers with the notion of terroir with different parent rocks considered to give birth to different wine styles. However, the relationship between wine properties and the underlying parent material raises a lot of controversy especially regarding the actual existence of rock-derived flavors in the wine (e.g. minerality). As far as the actual soil properties are concerned, the effect of soil physical properties is generally regarded as the most significant (e.g sandy soils being associated with lighter wines while those on clay with colored and tannic ones) mostly through control of water availability which ultimately modifies berry ripening conditions either directly by triggering biosynthetic pathways, or indirectly by altering vigor and yield components. The role of soil chemistry seems to be weakly associated to wine sensory characteristic, although N, K, S and Ca, but also soil pH, are often considered important in the overall soil effect.
Recently, in the light of evidence provided by precision agriculture studies reporting a high variability of vineyard soils, the spatial scale should also be taken into consideration in the evaluation of the soil effects on wines. While it is accepted that soil effects become more significant than climate on a local level, it is not clear whether these micro-variations of vineyard soils are determining in the terroir effect. Moreover, as terroir is not a set of only natural factors, the magnitude of the contribution of human-related factors (irrigation, fertilization, soil management) to the soil effect still remains ambiguous. Lastly, a major shortcoming of the majority of works about soil effects on wine characteristics is the absence of connection with actual vine physiological processes since all soil effects on grape and wine chemistry and sensorial properties are ultimately mediated through vine responses.
This article attempts to breakdown the main soil attributes involved in the terroir effect to suggest an improved understanding about soil’s true contribution to wine sensory characteristics. It is proposed that soil parameters per se are not as significant determining factors in the terroir effect but rather their mutual interactions as well as with other natural and human factors included in the terroir concept. Consequently, similarly to bioclimatic indices, composite soil indices (i.e. soil depth, water holding capacity, fertility, temperature etc), incorporating multiple soil parameters, might provide a more accurate and quantifiable means to assess the relative weight of the soil component in the terroir effect.

The plantation frame as a measure of adaptation to climate change

The mechanization of vineyard work originally led to a reduction in planting densities due to the lack of machinery adapted to the vineyard. The current availability of specific machinery makes it possible to establish higher planting densities. In this work, three planting densities (1.40×0.80 m, 1.80×1 m and 2.20×1.20 m, corresponding to 8928, 5555 and 3787 plants/ha respectively) were studied with four varieties autochthonous of Galicia (northwestern Spain): Albariño and Treixadura (white), Sousón and Mencía (red). The vines were trained in a vertical shoot positioning system using a single Royat cordon, and pruned to spurs with two buds each. Agronomic data (yield, pruning wood weight, Ravaz index) and oenological data in must were collected. The higher planting density (1.40×0.80 m) had no significant effect on grape yield per vine in white varieties, although production per hectare was much higher due to the greater number of plants. In red varieties, this planting density resulted in a significantly lower production per vine, compensated by the greater number of plants. In addition, it significantly reduced the Brix degree in the must of the Albariño, Treixadura and Sousón varieties, and increased the total acidity in the latter two and Mencía. It also caused an increase in extractable and total anthocyanins and IPT in red grapes. The effects of high planting density on grapes are of great interest for the adaptation of varieties in the context of climate change. In the future, it could be advisable to modify the limits imposed by the appellations of origin on the planting density of these varieties in order to obtain more balanced wines.

Impact of changes in pruning practices on vine growth and yield

A gradual decline in vineyards has been observed over the past twenty years worldwide. This might be explained by the climate change, practices change or the increase of dieback diseases. To increase the longevity of vines, we studied the impact of different pruning strategies in four adult and four young vineyards located in France and Spain. In France, vineyards were planted with Cabernet franc on 3309C while Spanish trials were planted with Tempranillo grafted on 110R. Vegetative expression, yield, quality of berries and wood vessels conductivity were measured. The distribution of vegetative expression, yield and berry composition between primary and secondary vegetation were quantified. Finally, tomography was used to evaluate the implication of the treatments on sap flows.
First results show that i) the respectful pruning leads to an increase of 30 to 50% more secondary shoots than the aggressive pruning in France and between 15 and 20% in Spain, ii) there is no major effect on the yield over the first two years following the implementation of the new pruning practices, although the proportion of clusters from suckers is higher on the respectful pruning method. On young vines, the development of the trunk according to a respectful pruning leads to a loss of harvest 2 years after planting. This is due to the removal, on the future trunk, of the green suckers which carrying bunches. This operation carried out in spring rather than during winter pruning, would promote a better leaf / fruit balance when the plant comes into production, and could lead to better hydraulic conduction in the vessels of the trunk. Maintaining these trials for several years will provide more robust data to assess the impact of these practices on the vines over the long term.

Impact of long term agroecological and conventional practices on subsurface soil microbiota in Macabeu and Xarel·lo vineyards

There is a growing trend on the transition from conventional to agroecological management of vineyards. However, the impact of practices, such as reduced-tillage, organic fertilization and cover crops, is not well-understood regarding the soil microbial diversity, and its relationship with the soil physicochemical properties in the subsurface depth near the rooting zone. Soil bacterial diversity is an important contributor towards plant health, productivity and response to environmental stresses. A field experiment was conducted by sampling subsurface soil bacterial community (NGS and qPCR) near to the root zone of Macabeu and Xarel·lo vineyards, located at the Penedes. 3 organic (ECO) and 3 conventional (CON) vineyards, with more than 10 years of respective management were sampled (n=5 each plot). ECO practices did not affect bacterial and fungal abundance but increased significantly the ammonium oxidizing bacteria and alpha-diversity (Inv.Simpson). Interestingly beta-diversity was significantly affected by the management strategy. ANOSIM-tests revealed a significative effect of the management (ecological vs conventional) and plot, on the soil microbial structure (ASV abundance). Main phyla depicted were Proteobacteria, Actinobacteria and Acidobacteria, whose relative abundances were not affected by the management. EdgeR assay revealed a significant increase of Cyanobacteria and decrease of Gemmatimonadetes and Firmicutes phyla in ECO. Interestingly, the grapevine variety was not correlated with the soil microbial community structure. Mantel-test revealed an important correlation (Spearman) of some physicochemical parameters with the soil microbiota structure, in order of importance: texture, EC, pH Ca/Mg, Mg/P, K+, Mg2+, Ca2+, SO42-, and OM. N-NH4 and NTK, which were higher in the ECO managed soils, did not correlated significantly with the soil microbiome population. The results revealed the importance of combining a deep physicochemical characterization of each replicate with the microbial diversity assessment to gain better insights on the relationship between soil microbiome and vineyard management.

Mobile device to induce heat-stress on grapevine berries

Studying heat stress response of grapevine berries in the field often relies on weather conditions during the growing season. We constructed a mobile heating device, able to induce controlled heat stress on grapes in vineyards. The heater consisted of six 150 W infrared lamps mounted in a profile frame. Heating power of the lamps could be controlled individually by a control unit consisting of a single board computer and six temperature sensors to reach a pre-set temperature. The heat energy applied to individual berries within a cluster decreases by the squared distance to the heat source, enabling the establishment of temperature profiles within individual clusters. These profiles can be measured by infrared thermography once a steady state has been reached. Radiant flux density received by a berry depending on the distance was calculated based on a view factor and measured lamp surface temperature and resulted to 665 Wm-2 at 7cm. Infrared thermography of the fruit surface was in good agreement with measurements conducted with a thermocouple inserted at epidermis level. In combination with infrared thermography, the presented device offers possibilities for a wide range of applications like phenotyping for heat tolerance in the field to proceed in the understanding of the complex response of plants to heat stress. Sunburn necrosis symptoms were artificially induced with the aid of the device for cv. Bacchus and cv. Sylvaner in the 2020 and 2021 growing season. Threshold temperatures for sunburn induction (LT5030min) were derived from temperature data of single berries and visual sunburn assessment, applying logistic regression. A comparison of threshold temperatures for the occurrence of sunburn necrosis confirmed the higher susceptibility of cv. Bacchus. The lower susceptibility of cv. Sylvaner did not seem to be related to its phenolic composition, rendering a thermoprotective role of berry phenolic compounds unlikely.

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IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Optimization and validation of a fully automated HS-SPME method for determination of VCCs and its application in wines submitted to accelerated ageing

Optimization and validation of a fully automated HS-SPME method for determination of VCCs and its application in wines submitted to accelerated ageing

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Publication date: June 23, 2022

Issue: IVAS 2022

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The rootstock, the neglected player in the scion transpiration even during the night

Water is the main limiting factor for yield in viticulture. Improving drought adaptation in viticulture will be an increasingly important issue under climate change. Genetic variability of water deficit responses in grapevine partly results from the rootstocks, making them an attractive and relevant mean to achieve adaptation without changing the scion genotype. The objective of this work was to characterize the rootstock effect on the diurnal regulation of scion transpiration. A large panel of 55 commercial genotypes were grafted onto Cabernet Sauvignon. Three biological repetitions per genotype were analyzed. Potted plants were phenotyped on a greenhouse balance platform capable of assessing real-time water use and maintaining a targeted water deficit intensity. After a 10 days well-watered baseline period, an increasing water deficit was applied for 10 days, followed by a stable water deficit stress for 7 days. Pruning weight, root and aerial dry weight and transpiration were recorded and the experiment was repeated during two years. Transpiration efficiency (ratio between aerial biomass and transpiration) was calculated and δ13C was measured in leaves for the baseline and stable water deficit periods. A large genetic variability was observed within the panel. The rootstock had a significant impact on nocturnal transpiration which was also strongly and positively correlated with maximum daytime transpiration. The correlations with growth and water use efficiency related traits will be discussed. Transpiration data were also related with VPD and soil water content demonstrating the influence of environmental conditions on transpiration. These results highlighted the role of the rootstock in modulating water deficit responses and give insights for rootstock breeding programs aimed at identifying drought tolerant rootstocks. It was also helpful to better define the mechanisms on which the drought tolerance in grapevine rootstocks is based on.

Permanent cover cropping with reduced tillage increased resiliency of wine grape vineyards to climate change

Majority of California’s vineyards rely on supplemental irrigation to overcome abiotic stressors. In the context of climate change, increases in growing season temperatures and crop evapotranspiration pose a risk to adaptation of viticulture to climate change. Vineyard cover crops may mitigate soil erosion and preserve water resources; but there is a lack of information on how they contribute to vineyard resiliency under tillage systems. The aim of this study was to identify the optimum combination of cover crop sand tillage without adversely affecting productivity while preserving plant water status. Two experiments in two contrasting climatic regions were conducted with two cover crops, including a permanent short stature grass (P. bulbosa hybrid), barley (Hordeum spp), and resident vegetation under till vs. no-till systems in a Ruby Cabernet (V. vinifera spp.) (Fresno) and a Cabernet Sauvingon (Napa) vineyard. Results indicated that permanent grass under no-till preserved plant available water until E-L stage 17. Consequently, net carbon assimilation of the permanent grass under no-till system was enhanced compared to those with barley and resident vegetation. On the other hand, the barley under no-till system reduced grapevine net carbon assimilation during berry ripening that led to lower content of nonstructural carbohydrates in shoots at dormancy. Components of yield and berry composition including flavonoid profile at either site were not adversely affected by factors studied. Switching to a permanent cover crop under a no-till system also provided a 9% and 3% benefit in cultural practices costs in Fresno and Napa, respectively. The results of this work provides fundamental information to growers in preserving resiliency of vineyard systems in hot and warm climate regions under context of climate change.

The effects of alternative herbicide free cover cropping systems on soil health, vine performance, berry quality and vineyard biodiversity in a climate change scenario in Switzerland

There is an urgent need in viticulture to adopt alternative herbicide-free soil management strategies to mitigate climate change, increase biodiversity, reduce plant protection products and improve soil quality while minimizing detrimental effects on grapevine’s stress tolerance and fruit quality. To propose sustainable solutions, adapted to different pedoclimatic conditions in Switzerland, we developed a multidisciplinary 4-year project, started in 2020. Objectives of the project are to a) evaluate the impact of green covers (spontaneous flora, winter cover crop and permanent ground cover) on environmental and agronomic parameters and b) develop subsequently innovative strategies for different viticultural contexts of Switzerland. The project is divided into 3 phases: 1) diagnosis, 2) on-farm and 3) on-station experiments. Phase 1) consisted in an assessment of 30 commercial vineyards all over Switzerland, where growers already use different herbicide-free soil management strategies. The most promising practices identified in this exploratory phase will be replicated in commercial vineyards across Switzerland (“on-farm”) as well as in a classical randomized block design in an experimental plot (“on-station”). For phase 1), measurements consisted in evaluation of soil status (compaction, structure, roots development), soil microbial diversity (metagenomics), plant diversity and biomass, vine physiology (water stress, vigor, leaf nitrogen) and berry quality (acidity, sugar, available nitrogen). Interestingly, the permanent ground cover resulted in a higher Shannon index thus a higher biodiversity as compared to the other itineraries. The winter cover crop increased vine nitrogen and vigor while deteriorating soil quality, leaving the soil more exposed and compacted likely due to more frequent tillage. The spontaneous flora led to higher berry sugar accumulation, less nitrogen and higher malic acid concentration putatively due to a higher water retention of the flora in a particularly wet vintage. Phases 2) and 3) are required to confirm those tendencies, over the 3 next vintages and different climatic conditions.

Climate change impacts: a multi-stress issue

With the aim of producing premium wines, it is admitted that moderate environmental stresses may contribute to the accumulation of compounds of interest in grapes. However the ongoing climate change, with the appearance of more limiting conditions of production is a major concern for the wine industry economic. Will it be possible to maintain the vineyards in place, to preserve the current grape varieties and how should we anticipate the adaptation measures to ensure the sustainability of vineyards? In this context, the question of the responses and adaptation of grapevine to abiotic stresses becomes a major scientific issue to tackle. An abiotic stress can be defined as the effect of a specific factor of the physico-chemical environment of the plants (temperature, availability of water and minerals, light, etc.) which reduces growth, and for a crop such as the vine, the yield, the composition of the fruits and the sustainability of the plants. Water stress is in many minds, but a systemic vision is essential for at least two reasons. The first reason is that in natural environments, a single factor is rarely limiting, and plants have to deal with a combination of constraints, as for example heat and drought, both in time and at a given time. The second reason is that plants, including grapevine, have central mechanisms of stress responses, as redox regulatory pathways, that play an important role in adaptation and survival. Here we will review the most recent studies dealing with this issue to provide a better understanding of the grapevine responses to a combination of environmental constraints and of the underlying regulatory pathways, which may be very helpful to design more adapted solutions to cope with climate change.

Investigating the impact of grape exposure and UV radiations on rotundone in Vitis vinifera L. Tardif grapes under field trial conditions

Rotundone is the main aroma compound responsible for peppery notes in wines whose biosynthesis is negatively affected by heat and drought. Through the alteration of precipitation regime and the increase in temperature during maturation, climate change is expected to affect wine peppery typicality. In this context there is a demand for developing sustainable viticultural strategies to enhance rotundone accumulation or limit its degradation. It was recently proposed that ultraviolet (UV) radiations could stimulate rotundone production. The aim of this study was to investigate under field trial conditions the impact of grape exposure and UV treatments on rotundone in Vitis vinifera L. Tardif, an almost extinct grape variety from south-west France that can express particularly high rotundone levels. Four different treatments were compared in 2021 to a control treatment using a randomised complete block design with three replications per treatment. Grape exposure was manipulated through early or late defoliation. Leaf and laterals shoots were removed at Eichorn Lorenz growth stages 32 or 34 on the morning-sun side of the canopy. During grape maturation, UV radiations were either reduced by 99% by installing UV radiation-shielding sheets, or applied four times using the Boxilumix™ non thermal device (Asclepios Tech, Tournefeuille) with the aim of activating plant signalling pathway. Loggers displayed in solar radiation shields were used to assess the effect of such shielding sheets on air temperature within the bunch zone. The composition of grapes subjected to these treatments will be soon analysed for their rotundone content and basic classical laboratory analyses. Grapes will be harvested to elaborate wines under standardized small-scale vinification conditions (60kg) that will be assessed by a trained sensory panel.