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…

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…).

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.

Better understand the soil wet bulb formation with subsurface or aerial drip irrigation in viticulture

The gradual change in rainfall patterns experienced in the south of France vineyards, especially around the Mediterranean sea, means that the vines are increasingly subject to summer drought. The winegrowers developped the use of irrigation techniques to ensure the maintenance of competitive yields in the production of wines under Protected Geographical Indication label. In practice, drip irrigation pipes can be installed above the ground or buried into the soil as well as at different distances from the vine row. The objective of this study was to examine the profiles of the wet bulbs of the soil obtained from two drip irrigation systems : aerial drip located under the vine row and subsurface drip placed in the middle of the inter-row. This experiment took place over two consecutive seasons (2020-2021) on a 3.4 ha Viognier plot in the Mediterranean region (PGI Oc, France) on sandy clay soil. The annual rainfalls were less than 400 mm. Soil water content probes were installed at different depths (20 – 40 – 60 – 80 cm) and at different lateralities from the vine row (30 – 60 – 90 – 120 cm) to control the formation of the soil wet bulb during irrigation. The mapping and the analysis of the data allowed a better understanding and differentiation of the water percolation when irrigating with subsurface or aerial drip. For the same amount of water and without differences of vine water status, it is shown that in a subsurface drip irrigation situation, the size of the wet bulb formed is larger than in aerial drip irrigation system.

Grapevine xylem embolism resistance spectrum reveals which varieties have a lower mortality risk in a future dry climate

Wine growing regions have recently faced intense and frequent droughts that have led to substantial economical losses, and the maintenance of grapevine productivity under warmer and drier climate will rely notably on planting drought-resistant cultivars. Given that plant growth and yield depend on water transport efficiency and maintenance of photosynthesis, thus on the preservation of the vascular system integrity during drought, a better understanding of drought-related hydraulic traits that have a significant impact on physiological processes is urgently needed. We have worked towards this end by assessing vulnerability to xylem embolism in 30 grapevine commercial varieties encompassing red and white Vitis vinifera varieties, hybrid varieties characterized by a polygenic resistance for powdery and downy mildew, and commonly used rootstocks. These analyses further allowed a global assessment of wine regions with respect to their varietal diversity and resulting vulnerability to stem embolism. Hybrid cultivars displayed the highest vulnerability to embolism, while rootstocks showed the greatest resistance. Significant variability also arose among Vitis vinifera varieties, with Ψ12 and Ψ50 values ranging from -0.4 to -2.7 MPa and from -1.8 to -3.4 MPa, respectively. Cabernet franc, Chardonnay and Ugni blanc featured among the most vulnerable varieties while Pinot noir, Merlot and Cabernet Sauvignon ranked among the most resistant. In consequence, wine regions bearing a significant proportion of vulnerable varieties, such as Poitou-Charentes, France and Marlborough, New Zealand, turned out to be at greater risk under drought. These results highlight that grapevine varieties may not respond equally to warmer and drier conditions, outlining the importance to consider hydraulic traits associated with plant drought tolerance into breeding programmes and modeling simulations of grapevine yield maintenance under severe drought. They finally represent a step forward to advise the wine industry about which varieties and regions would have the lowest risk of drought-induced mortality under climate change.

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.