WAC 2022 banner
IVES 9 IVES Conference Series 9 WAC 9 WAC 2022 9 3 - WAC - Oral 9 Impact of aspects of the polysaccharide structure of mannoproteins on their interactions with Enological Tannins

Impact of aspects of the polysaccharide structure of mannoproteins on their interactions with Enological Tannins

Abstract

Mannoproteins (MPs) with different structure of their polysaccharide part (branching, substitutions, …) were used to better understand the impact of characteristics of the usual structure of MPs when interacting with Grape Seed Tannins (ST). 

From four Saccharomyces cerevisiae strains we obtain four MP pools: an enological strain LMD47 (presenting high levels of N-glycosylation and O-Mannosylation), a wild-type BY4742 strain (used as reference), and its mutants ΔMnn4 (with no mannosyl-phosphorylation) and ΔMnn2 (with a linear N-glycosylation backbone). The extraction method applied, with the exclusive enzymatic activity of Endo-β-1,3-Glucanase of Trichoderma sp. (E-LAMSE, Megazym), preserved the indigenous structure of mannoproteins to their utmost extent. Characterizations of the pools confirmed differences among the polysaccharide moieties of the four MPs regarding charge, mannose/glucose ratio, and branching degrees but no differences between their protein moieties.

The formation and evolution of colloidal aggregates due to interactions between MPs and ST at different concentrations were evaluated through Dynamic Light Scattering (DLS), while the number of colloidal aggregates formed and the particle size distribution were assessed by Nanoparticle Tracking Analysis (NTA). The possible differences in the mechanisms of interaction among the four kinds of mannoproteins were analyzed through Isothermal Titration Calorimetry (ITC).

DLS and NTA experiments indicated an immediate formation of colloidal aggregates, in which the final particle size and concentration were dependent on the ST/MP ratio. Whenever the latter was extremely high, a very progressive flocculation related to a reversible aggregation occurred. The kinetics of this instability phenomenon was dependent on the polysaccharide structure of MPs. ITC analysis showed two different kinds of interactions: an intense exothermic one susceptible to temperature, and a much weaker interaction (as for enthalpy release) less thermo-dependent, possibly related to H-bonding and hydrophobic interactions, respectively. 

Neither the absence of mannosyl phosphate groups, the absence of ramifications on the outer chains of the N-glycosylation, nor the protein glycosylation overexpression seem to play a decisive role in those interactions. However, these structural differences affected the stability of MP-ST colloids formed at specific concentrations and slightly changed the enthalpy exchange profiles.

DOI:

Publication date: June 13, 2022

Issue: WAC 2022

Type: Article

Authors

Assunção Bicca, Céline, Poncet-Legrand, Julie, Mekoue Nguela, Thierry, Doco, Aude, Vernhet

Presenting author

Assunção Bicca – Université de Montpellier

Unité Mixte de Recherche Sciences Pour l’OEnologie, Institut Agro, INRAE, Université de Montpellier, Montpellier, France | Lallemand SAS | Unité Mixte de Recherche Sciences Pour l’OEnologie

Contact the author

Keywords

Mannoproteins – Colloidal Stability – Polysaccharide/Polyphenol Interactions – Wine macromolecules

Tags

IVES Conference Series | WAC 2022

Citation

Related articles…

Grape must quality and mesoclimatic variability in Fruška Gora wine-growing region, Serbia

The Fruška Gora mountain is a traditional wine-growing region in Serbia situated in the Pannonian Basin. Due to such a position, the vicinity of the Danube River and the presence of concave configuration, it is suitable for grape production. This paper provides analyses of spatial variations in meteorological parameters and grape juice quality within Fruška Gora wine region over three consecutive vintages (2018-2020). The examined period can be defined as warm with cool nights during September (AVG 18,9°C; GDD 1918°C; CI 12°CF) and with the presence of mesoclimatic variability. The East part of the study area was somewhat drier and hotter compared to other parts of the region. The analyses of grape must samples (190 in total) of five cultivars (Cabernet-Sauvignon, Merlot, Chardonnay, Sauvignon blanc and Grašac (Welschriesling)) commonly grown across the region (19 sites), were performed using Fourier Transform Infrared Technology (FTIR). Among all cultivars, Sauvignon blanc was harvested first in the East area (DOY=246±5, GDD at harvest=1552±74, 22.2±0.7 °Brix), while the latest harvest was recorded for Cabernet-Sauvignon in the West (DOY=283±5, GDD at harvest=1936±187, 23.4±1.0 °Brix ). Both the red and white cultivars had higher acidity and YAN in the grape must if the vines were grown in the North and East compared to South and West areas. According to PCA analysis, Grašac showed the lowest variation in grape must chemical composition. Thus, the results confirm that Grašac is the most stable cultivar in Fruška Gora. All monitored cultivars reached technological fruit ripeness by the end of the growing season. However, it was difficult to reach full ripeness of red cultivars, mostly beacuse of uncoupling of technolocical and phenolic ripeness. Thus, Cabernet-Sauvignon had higher variations in GDD sums at harvest compared to other cultivars, which probably increased variations in grape must quality.

How can historical cultivars mitigate the effects of climate change?

IFV, INRAe and the national network “Partenaires de la Sélection Vigne” representing 37 organizations from the different wine regions, have been working increasingly closely over the last 2 decades towards the preservation of the French varietal patrimony. There are approximately 600 patrimonial varieties according to INRAe and SupAgro Montpellier experts, including ancient cultivars (400) and intravarietal crossbreeds obtained since the 19th century. In the context of a drastic reduction in such varieties from the mid 1980’s in favor of mainstream varieties, it was essential to carry out an inventory of old vines and vineyards. INRAe Vassal collection plays a key role here as it holds the largest diversity available, along with a rich bibliography and herbariums, offering us the opportunity to document and double check the identity of a cultivar, consolidating the expertise of ampelographers. The work is carried out in several stages, from verifying the existence of a variety in a small region, through to rehabilitation. During this session, the authors present the process that leads to the official registration of a variety. After this, IFV selection center takes over to initiate the process of selection and propagation. A specific focus within regions such as the Alps, Champagne and the South-West will provide details of the full procedure. Bia, Bouysselet, Chardonnay rose, Mecle and the aptly named Tardif, are some of the cultivars that have followed this procedure. Furthermore, a recent regulation established by INAO on “varieties of interest for adaptation purposes” might boost uptake by growers. Since 2006, 36 historical cultivars have been registered. Most of these have been neglected in the past due to late maturity, lack of sugar and high titratable acidity at harvest time. Such characteristics are today considered as positive qualities, not only in mitigation of the effects of climate change, but also as an opportunity for restoring diversity…

20-Year-Old data set: scion x rootstock x climate, relationships. Effects on phenology and sugar dynamics

Global warming is one of the biggest environmental, social, and economic threats. In the Douro Valley, change to the climate are expected in the coming years, namely an increase in average temperature and a decrease in annual precipitation. Since vine cultivation is extremely vulnerable and influenced by the climate, these changes are likely to have negative effects on the production and quality of wine.
Adaptation is a major challenge facing the viticulture sector where the choice of plant material plays an important role, particularly the rootstock as it is a driver for adaptation with a wide range of effects, the most important being phylloxera, nematode and salt, tolerance to drought and a complex set of interactions in the grafted plant.
In an experimental vineyard, established in the Douro Region in 1997, with four randomized blocs, with five varieties, Touriga Nacional, Tinta Barroca, Touriga Franca and Tinta Roriz, grafted in four rootstocks, Rupestris du Lot, R110, 196-17C, R99 and 1103P, data was collected consecutively over 20 years (2001-2020). Phenological observations were made two to three times a week, following established criteria, to determine the average dates of budbreak, flowering and veraison. During maturation, weekly berry samples were taken to study the dynamics of sugar accumulation, amongst other parameters. Climate data was collected from a weather station located near the vineyard parcel, with data classified through several climatic indices.
The results achieved show a very low coefficient of variations in the average date of the phenophases and an important contribution from the rootstock in the dynamic of the phenology, allowing a delay in the cycle of up to10-12 days for the different combinations. The Principal Component Analysis performed, evaluating trends in the physical-chemical parameters, highlighted the effect of the climate and rootstock on fruit quality by grape varieties.

Late frost protection in Champagne

Probably one of the most counterintuitive impacts of climate change on vine is the increased frequency of late frost. Champagne, due to its septentrional position is historically and regularly affected by this meteorological hazard. Champagne has therefore developed a strong experience in frost protection with first experiments dating from the end of 19th century. Frost protection can be divided in two parts: passive and active. Passive protection includes all the methods that do not seek to modify the vine’s environment or resistance at the time of frost. The most iconic passive protection in Champagne is the establishment of the individual reserve. This reserve allows to stock a certain quantity of clear wine during a surplus year to compensate a meteorological hazard like frost during the following years. Other common passive methods are the control of planting area (walls, bushes, topography), the choice of grape variety, late pruning, or the impact of grass cover and tillage. Active frost protection is also divided in two parts. Most of the existing techniques tend to modify vine’s environment. Most of the time they provide warmth (candles, heaters, windmills, heating cables…), or stabilise bud’s temperature above a lethal threshold (water sprinkling). The other way to actively fight is to enhance the resistance of buds to frost (elicitors). The Comité Champagne evaluates frost protection methods following three main axes: the efficiency, the profitability, and the environmental impact through a lifecycle assessment. This study will present the results on both passive and active protection following these three axes.

The potential of multispectral/hyperspectral technologies for early detection of “flavescence dorée” in a Portuguese vineyard

“Flavescence dorée” (FD) is a grapevine quarantine disease associated with phytoplasmas and transmitted to healthy plants by insect vectors, mainly Scaphoideus titanus. Infected plants usually develop symptoms of stunted growth, unripe cane wood, leaf rolling, leaf yellowing or reddening, and shrivelled berries. Since plants can remain symptomless up to four years, they may act as reservoirs of FD contributing to the spread of the disease. So far, conventional management strategies rely mainly on the insecticide treatments, uprooting of infected plants and use of phytoplasma-free propagation material. However, these strategies are costly and could have undesirable environmental impacts. Thus, the development of sustainable and noninvasive approaches for early detection of FD and its management are of great importance to reduce disease spread and select the best cultural practices and treatments. The present study aimed to evaluate if multispectral/hyperspectral technologies can be used to detect FD before the appearance of the first symptoms and if infected grapevines display a spectral imaging fingerprint. To that end, physiological parameters (leaf area, chlorophyll content and photosynthetic rate) were collected in concomitance to the measurements of plant reflectance (using both a portable apparatus and a remote sensing drone). Measurements were performed in two leaves of 8 healthy and 8 FD-infected grapevines, at four timepoints: before the development of disease symptoms (21st June); and after symptoms appearance (ii) at veraison (2nd August); at post-veraison (11th September); and at harvest (25th September). At all timepoints, FD infected plants revealed a significant decrease in the studied physiological parameters, with a positive correlation with drone imaging data and portable apparatus analyses. Moreover, spectra of either drone imaging and portable apparatus showed clear differences between healthy and FD-infected grapevines, validating multispectral/ hyperspectral technology as a potential tool for the early detection of FD or other grapevine-associated diseases.