OENO IVAS 2019 banner
IVES 9 IVES Conference Series 9 OENO IVAS 9 OENO IVAS 2019 9 Chemical and Biochemical reactions, including grape and wines microorganisms impact 9 Oenological tannins to prevent Botrytis cinerea damage: reduction of laccase activity

Oenological tannins to prevent Botrytis cinerea damage: reduction of laccase activity

Abstract

Oenological tannins are classified as hydrolysable and condensed tannins. Their use in winemaking is only authorized, to facilitate wine fining. Nevertheless, tannins could also be used to prevent laccase effects. 

Indeed, our group has recently proved their effects against laccase damage [1]. The goal of this study was to better understand the mechanism of action of oenological tannins on laccase activity induced by Botrytis cinerea. Five oenological tannins were used (gallotannin, ellagitannin, quebracho, grape-skin and grape-seed) and compared with ascorbic acid (AA) and sulfur dioxide (SO2). Oenological tannins, AA and SO2 were added to botrytized must at different doses. After 4 minutes, laccase activity was measured by the syringaldazine method [2] using different concentration of subtract. Enzymatic kinetic constants (Km/Vmax) were determined according to Michaelis-Menten model. Furthermore, B. cinerea (strain 213) was grown in a stimulating liquid medium for laccase production [3]. The molecular weight (MW) and the effect of bentonite and tannins upon laccase were studied by SDS-PAGE. The results confirm that all oenological tannins inhibit laccase activity and that the higher the dose the lower the laccase activity. In this way, gallotannin, grape-seed and skins tannins seem to be the most effective tannins. All the tannins were as effective as AA, even though SO2 was clearly the most effective inhibitor. The laccase produced by B. cinerea had a MW of 95 kDa. After bentonite treatment all wine protein bands disappeared and laccase band decreased slightly although its activity remains stable. The interaction between different oenological tannins and laccase was analyzed by measuring the reduction of the intensity of the laccase band. In general, the reduction of band intensity correlates with the reduction of laccase enzymatic activity. 

It can be concluded therefore that oenological tannins a good candidate to prevent laccase effects, helping to diminish the SO2 dose in grapes infected by B. cinerea. 

[1] Vignault, A.; Pascual, O.; Jourdes, M.; Moine, V.; Fermaud, M.; Roudet, J.; Canals, J.M.; Teissedre, P-L.; Zamora, F. Oeno One, 2019. 
[2] Urbano Cuadrado, M.; Pérez-Juan, P.M.; Luque de Castro, M.; Gomez-Nieto, M.A., Anal Chim Acta, 2005, 553, 99-104. 
[3] Quijada-Morin, N.; Garcia, F.; Lambert, K.; Walker, A.S.; Tiers, L.; Viaud, M.; Sauvage, F-X.; Hirtz, C.; Saucier, C.; Aust J Grape Wine Res, 2018, 24, 241-251.

DOI:

Publication date: June 19, 2020

Issue: OENO IVAS 2019

Type: Article

Authors

Fernando Zamora (1), Adeline Vignault (2), Jordi Gombau (1), Michael Jourdes (2), Virginie Moine (3), Joan Miquel Canals (1), Pierre-Louis Teissedre (2)

1) Departament de Bioquímica i Biotecnologia. Facultat d’Enologia. Universitat Rovira i Virgili. C/ Marcel.lí Domingo 1, 43007-Tarragona (Spain)
2) Unitéde recherche Oenologie, EA 4577, USC 1366 INRA, ISVV, Universitéde Bordeaux, Bordeaux INP, F33882 Villenave d’Ornon France
3) Laffort, 11 rue Aristide berges, 33270 Floirac, France

Contact the author

Keywords

Oenological tannins, Botrytis cinerea laccase, Kinetics, SDS-PAGE 

Tags

IVES Conference Series | OENO IVAS 2019

Citation

Related articles…

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.

Using δ13C and hydroscapes as a tool for discriminating cultivar specific drought response

Measurement of carbon isotope discrimination in berry juice sugars at maturity (δ13C) provides an integrated assessment of water use efficiency (WUE) during the period of berry ripening, and when collected over multiple seasons can be used as an indication of drought stress response. Berry juice δ13C measurements were carried out on 48 different varieties planted in a common garden experiment in Bordeaux, France from 2014 through 2021 and were paired with midday and predawn leaf water potential measurements on the same vines in a subset of six varieties. The aim was to discriminate a large panel of varieties based on their stomatal behaviour and potentially identify hydraulic traits characterizing drought tolerance by comparing δ13C and hydroscapes (the visualisation of plant stomatal behaviour as a response to predawn water potential). Cluster analysis found that δ13C values are likely affected by the differing phenology of each variety, resulting in berry ripening of different varieties taking place under different stress conditions within the same year. We accounted for these phenological differences and found that cluster analysis based on specific δ13C metrics created a classification of varieties that corresponds well to our current empirical understanding of their relative drought tolerances. In addition, we analysed the water potential regulation of the subset of six varieties (using the hydroscape approach) and found that it was well correlated with some δ13C metrics. Surprisingly, a variety’s water potential regulation (specifically its minimum critical leaf water potential under water deficit) was strongly correlated to δ13C values under well-watered conditions, suggesting that base WUE may have a stronger impact on drought tolerance than WUE under water deficit. These results give strong insights on the innate WUE of a very large panel of varieties and suggest that studies of drought tolerance should include traits expressed under non-limiting conditions.

Grapevine yield estimation in a context of climate change: the GraY model

Grapevine yield is a key indicator to assess the impacts of climate change and the relevance of adaptation strategies in a vineyard landscape. At this scale, a yield model should use a number of parameters and input data in relation to the information available and be able to reproduce vineyard management decisions (e.g. soil and canopy management, irrigation). In this study, we used data from six experimental sites in Southern France (cv. Syrah) to calibrate a model of grapevine yield limited by water constraint (GraY). Each yield component (bud fertility, number of berries per bunch, berry weight) was calculated as a function of the soil water availability simulated by the WaLIS water balance model at critical phenological phases. The model was then evaluated in 10 grapegrowers’ plots, covering a diversity of biophysical and technical contexts (soil type, canopy size, irrigation, cover crop). We identified three critical periods for yield formation: after flowering on the previous year for the number of bunches and berries, around pre-veraison and post-veraison of the same year for mean berry weight. Yields were simulated with a model efficiency (EF) of 0.62 (NRMSE = 0.28). Bud fertility and number of berries per bunch were more accurately simulated (EF = 0.90 and 0.77, NRMSE = 0.06 and 0.10, respectively) than berry weight (EF = -0.31, NRMSE = 0.17). Model efficiency on the on-farm plots reached 0.71 (NRMSE = 0.37) simulating yields from 1 to 8 kg/plant. The GraY model is an original model estimating grapevine yield evolution on the basis of water availability under future climatic conditions.  It allows to evaluate the effects of various adaptation levers such as planting density, cover crop management, fruit/leaf ratio, shading and irrigation, in various production contexts.

Measurement of redox potential as a new analytical winegrowing tool

Excell laboratory has initiated the development of an analytical method based on electrochemistry to evaluate the ability of wines to undergo or resist to oxidative phenomena. Electrochemistry is a powerful tool to probe reactions involving electron transfers and offers possibility of real-time measurements. In that context, the laboratory has implemented electrochemical analysis to assess oxidation state of different wine matrices but also in order to evaluate oxidative or reduced character of leaf and soil. Initially, our laboratory focused on dosage of compounds involved in responses of plant stresses and we were also interested in microbiological activity of soils. These analyses were compared with the measurement of redox potential (Eh) and pH which are two fundamental variables involved in the modulation of plant metabolism. Indeed, the variation of redox states of the plant reflects its biological activity but also its capacity to absorb nutriments. The Eh-pH conditions mainly determine metabolic processes involved in soil and leaf and our goal is to determine if this combined analytical approach will be sufficiently precise to detect biological evolutions (plant health, parasitic attack…).

Optimizing stomatal traits for future climates

Stomatal traits determine grapevine water use, carbon supply, and water stress, which directly impact yield and berry chemistry. Breeding for stomatal traits has the strong potential to improve grapevine performance under future, drier conditions, but the trait values that breeders should target are unknown. We used a functional-structural plant model developed for grapevine (HydroShoot) to determine how stomatal traits impact canopy gas exchange, water potential, and temperature under historical and future conditions in high-quality and hot-climate California wine regions (Napa and the Central Valley). Historical climate (1990-2010) was collected from weather stations and future climate (2079-99) was projected from 4 representative climate models for California, assuming medium- and high-emissions (RCP 4.5 and 8.5). Five trait parameterizations, representing mean and extreme values for the maximum stomatal conductance (gmax) and leaf water potential threshold for stomatal closure (Ψsc), were defined from meta-analyses. Compared to mean trait values, the water-spending extremes (highest gmax or most negative Ysc) had negligible benefits for carbon gain and canopy cooling, but exacerbated vine water use and stress, for both sites and climate scenarios. These traits increased cumulative transpiration by 8 – 17%, changed cumulative carbon gain by -4 – 3%, and reduced minimum water potentials by 10 – 18%. Conversely, the water-saving extremes (lowest gmax or least negative Ψsc) strongly reduced water use and stress, but potentially compromised the carbon supply for ripening. Under RCP 8.5 conditions, these traits reduced transpiration by 22 – 35% and carbon gain by 9 – 16% and increased minimum water potentials by 20 – 28%, compared to mean values. Overall, selecting for more water-saving stomatal traits could improve water-use efficiency and avoid the detrimental effects of highly negative canopy water potentials on yield and quality, but more work is needed to evaluate whether these benefits outweigh the consequences of minor declines in carbon gain for fruit production.