OENO IVAS 2019 banner
IVES 9 IVES Conference Series 9 Importance of matrix effects (wine composition) on protein stability tests of white and rosé wines

Importance of matrix effects (wine composition) on protein stability tests of white and rosé wines

Abstract

The presence of unstable proteins in wines can affect their stability and clarity. Before bottling, winemakers need to be sure that the wine is stable. A large number of stability tests have been proposed, usually based on heating a sample with a specific time-temperature couple. In practice, none is effective to accurately assess the risk of instability. Moreover, the interpretation of the results of these tests changes according to the region. 

The aim of this work is to compare, on 55 wines (4 vintages, 7 varieties, 5 areas), the most common heat test (30 minutes at 80°C) with the turbidity measured after 15 days at 35 °C on bottled wines. Proteins were analyzed in 33 cases. In addition, 10 wines were heated at 40 °C/30 min, 40°C/4 hours, 35 °C/15 days and 80 °C/30 min and the residual proteins analyzed. 

The results show no correlation between turbidity after heat test 80 °C/30 min and after 15 days at 35 °C. For some wines, especially Gewurztraminer ones, turbidity after heating at 80 °C can reach 330 NTU without any visual haze at 35 °C (< 3 NTU). Similar results are obtained when the heat test is performed after adjustment of pH to 3.4. The turbidity after heat test 80°C/30 min increases with pH, particularly above 3.6, which is not so unusual for Gewurztraminer wines. The pH effect is less significant at 40 °C. Finally, pH values alone cannot explain the different behaviors of wines. 

On the other hand, protein composition in wines depends on their pH. Thaumatin Like proteins (TL) 19 kDa, TL 22kDa and Invertases are present in almost all wines. Half of them contains Lipid Transfer Protein (LTP) and only a few Chitinases and β-Glucanase. These proteins are present when pH is lower than 3.5, probably because low pH favor Chitinase and-glucanase conformational changes and precipitation. 

Protein analysis after heating these various wines at different time-temperature couples led to this ranking: 
Chitinases are sensitive at low temperature (40 °C) and resist better at pH 3.7; 
TL 22kDa are sensitive, especially in Rosé wines; 
TL 19kDa are more stable, but their sensitivity depends on the pH; 
Invertase unfold between 60 and 80°C but is not affected by the pH; 
LTP can resist up to 80 °C. 

Turbidity after usual heat test 80 °C/30 min increases with total proteins concentration and pH. This is not observed after 15 days at 35 °C or 4 hours at 40 °C. These tests may be better to evaluate the actual risk of instability after bottling.

Related articles…

Anthocyanin profile is differentially affected by high temperature, elevated CO2 and water deficit in Tempranillo (Vitis vinifera L.) clones

Anthocyanin potential of grape berries is an important quality factor in wine production. Anthocyanin concentration and profile differ among varieties but it also depends on the environmental conditions, which are expected to be greatly modified by climate change in the future. These modifications may significantly modify the biochemical composition of berries at harvest, and thus wine typicity. Among the diverse approaches proposed to reduce the potential negative effects that climate change may have on grape quality, genetic diversity among clones can represent a source of potential candidates to select better adapted plant material for future climatic conditions. The effects of individual and combined factors associated to climate change (increase of temperature, rise of air CO2 concentration and water deficit) on the anthocyanin profile of different clones of Tempranillo that differ in the length of their reproductive cycle were studied. The aim was to highlight those clones more adapted to maintain specific Tempranillo typicity in the future. Fruit-bearing cuttings were grown in controlled conditions under two temperatures (ambient temperature versus ambient temperature + 4ºC), two CO2 levels (400 ppm versus 700 ppm) and two water regimes (well-watered versus water deficit), both in combination or independently, in order to simulate future climate change scenarios. Elevated temperature increased anthocyanin acylation, whereas elevated CO2 and water deficit favoured the accumulation of malvidin derivatives, as well as the acylation and tri-hydroxylation level of anthocyanins. Although the changes in anthocyanin profile observed followed a common pattern among clones, such impact of environmental conditions was especially noticeable in one of the most widely distributed Tempranillo clones, the accession RJ43.

What are the optimal ranges and thresholds for berry solar radiation for flavonoid biosynthesis?

In wine grape production, canopy management practices are applied to control the source-sink balance and improve the cluster microclimate to enhance berry composition. The aim of this study was to identify the optimal ranges of berry solar radiation exposure (exposure) for upregulation of flavonoid biosynthesis and thresholds for their degradation, to evaluate how canopy management practices such as leaf removal, shoot thinning, and a combination of both affect the grapevine (Vitis vinifera L. cv. Cabernet Sauvignon) yield components, berry composition, and flavonoid profile under context of climate change. First experiment assessed changes in the grape flavonoid content driven by four degrees of exposure. In the second experiment, individual grape berries subjected to different exposures were collected from two cultivars (Cabernet Sauvignon and Petit Verdot). The third experiment consisted of an experiment with three canopy management treatments (i) LR (removal of 5 to 6 basal leaves), (ii) ST (thinned to 24 shoots per vine), and (iii) LRST (a combination of LR and ST) and an untreated control (UNT). Berry composition, flavonoid content and profiles, and 3-isobutyl 2-methoxypyrazine were monitored during berry ripening. Although increasing canopy porosity through canopy management practices can be helpful for other purposes, this may not be the case of flavonoid compounds when a certain proportion of kaempferol was achieved. Our results revealed different sensitivities to degradation within the flavonoid groups, flavonols being the only monitored group that was upregulated by solar radiation. Within different canopy management practices, the main effects were due to the ST. Under environmental conditions given in this trial, ST and LRST hastened fruit maturity; however, a clear improvement of the flavonoid compounds (i.e., greater anthocyanin) was not observed at harvest. Methoxypyrazine berry content decreased with canopy management practices studied. Although some berry traits were improved (i.e. 2.5° Brix increase in berry total soluble solids) due to canopy management practices (ST), this resulted in a four-fold increase in labor operations cost, two-fold decrease in yield with a 10-fold increase in anthocyanin production cost per hectare that should be assessed together as the climate continues to get hot.

Estimating bulk stomatal conductance of grapevine canopies

In response to changes in their environment, grapevines regulate transpiration using various physiological mechanisms that alter conductance of water through the soil-plant-atmosphere continuum. Expressed as bulk stomatal conductance at the canopy scale, it varies diurnally in response to changes in vapor pressure deficit and net radiation, and over the season to changes in soil water deficits and hydraulic conductivity of both soil and plant. It is necessary to characterize the response of conductance to these variables to better model how vine transpiration also responds to these variables. Furthermore, to be relevant for vineyard-scale modeling, conductance is best characterized using data collected in a vineyard setting. Applying a crop canopy energy flux model developed by Shuttleworth and Wallace, bulk stomatal conductance was estimated using measurements of individual vine sap flow, temperature and humidity within the vine canopy, and estimates of net radiation absorbed by the vine canopy. These measurements were taken on several vines in a non-irrigated vineyard in Bordeaux France, using equipment that did not interfere with ongoing vineyard operations. An inverted Penman-Monteith equation was then used to calculate bulk stomatal conductance on 15-minute intervals from July to mid-September 2020. Time-series plots show significant diurnal variation and seasonal decreases in conductance, with overall values similar to those in the literature. Global sensitivity analysis using non-parametric regression found transpiration flux and vapor pressure deficit to be the most important input variables to the calculation of bulk stomatal conductance, with absorbed net radiation and bulk boundary layer conductance being much less important. Conversely, bulk stomatal conductance was one of the most important inputs when calculating vine transpiration, further emphasizing the need for characterizing its response to environmental changes for use in vineyard water use modeling.

Low-cost sensors as a support tool to monitor soil-plant heat exchanges in a Mediterranean vineyard

Mediterranean viticulture is increasingly exposed to more frequent extreme conditions such as heat waves. These extreme events co-occur with low soil water content, high air vapor pressure deficit and high solar radiant energy fluxes and result in leaf and berry sunburn, lower yield, and berry quality, which is a major constraint for the sustainability of the sector. Grape growers must find ways to proper and effectively manage heat waves and extreme canopy and berry temperatures. Irrigation to keep soil moisture levels and enable adequate plant turgor, and convective and evaporative cooling emerged as a key tool to overcome this major challenge. The effects of irrigation on soil and plant water status are easily quantifiable but the impact of irrigation on soil and canopy temperature and on heat convection from soil to cluster zone remain less characterized. Therefore, a more detailed quantification of vineyard heat fluxes is highly relevant to better understand and implement strategies to limit the effects of extreme weather events on grapevine leaf and berry physiology and vineyards performance. Low-cost sensor technologies emerge as an opportunity to improve monitoring and support decision making in viticulture. However, validation of low-cost sensors is mandatory for practical applicability. A two-year study was carried in a vineyard in Alentejo, south of Portugal, using low-cost thermal cameras (FLIR One, 80×60 pixels and FLIR C5, 160×120 pixels, 8-14 µm, FLIR systems, USA) and pocket thermohygrometers (Extech RHT30, EXTECH instruments, USA) to monitor grapevine and soil temperatures. Preliminary results show that low-cost cameras can detect severe water stress and support the evaluation of vertical canopy temperature variability, providing information on soil surface temperature. All these thermal parameters can be relevant for soil and crop management and be used in decision support systems.

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).