Development, validation and application of a fast UHPLC-HRMS method for the analysis of amino acids and biogenic amines in wines and musts.

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.

In viticulture, the challenges of local climate modelling are multiple: taking into account the local environment, fine temporal and spatial scales, reliable time series of climate data, ease of implementation and reproducibility of the method. At the local scale, recent studies have demonstrated the contribution of spatialization methods for ground-based climate observation data considering topographic factors such as altitude, slope, aspect, and geographic coordinates (Le Roux et al, 2017; De Rességuier et al, 2020). However, these studies have shown questions in terms of the reproducibility and sustainability of this type of climate study. In this context, we evaluated the potential of MODIS thermal satellite images validated with ground-based climate data (Morin et al, 2020). Previous studies have been encouraging, but questions remain to be explored at the regional scale, particularly in the dynamics of the massive use of bioclimatic indices to classify the climate of wine regions. The results at the local scale were encouraging, but this approach was tested in the current study at the regional scale. Several objectives were set: 1) to evaluate the downscaling method for land surface temperature time series, 2) to identify regional thermal structure variations. We used weekly minimum and maximum surface temperature time series acquired by MODIS satellites at a spatial resolution of 1000 m and downscaled at 500 m using topographical variables. Two types of analyses were performed:

In the study of terroir, a separate analysis of its many component factors can be of great help in accurately identifying a vineyard’s natural elements that impact wine quality and typicity. This research used a dedicated pluri-disciplinary approach to investigate the ecological characteristics, including geology and geographical features, of 14 vineyards that produce Gewürztraminer and Sauvignon Blanc cultivars in the alpine Alto Adige DOC wine region. Both the geopedological method using Vineyards Geological Identity (VGI) and the new Solar Radiaton Identity (SRI) topoclimatic classification method were used to provide analytical measurements and qualitative/quantitative characterisations. In addition, wide-ranging targeted and untargeted oenological and chemical analyses were carried out on grapes, musts and wines to correlate the soils’ geomineral and physical conditions with the biochemical properties of their fruits and wines. The research identified strong correlations between vineyard geo-identity and wine biofingerprint, confirming a mineral traceability of strontium rubidium ratio and some minerals distinctive to the local geology, such as K, Ca, Ag, Ba and Mn.  The study also discovered that particular geomineral and physical soil conditions of the studied vineyards are related to the different amount of amino acids, primary varietal aromas and polyphenols found in grapes, musts and wines. The research confirmed that winemaking technologies support oenological quality, although in some cases, human practices can overpower certain characteristic elements in wine, erasing the typical imprint left by the vineyards’ natural terroir, which becomes less traceable. Terroir abiotic ecological factors and vineyard identity can be classified in detail using the new VGI and SRI analysis methods to discover interrelationships between geo-pedological and topoclimatic conditions that impact wine quality. These methods are also helpful in identifying which ecological elements are exclusive to a particular vineyard or wine sub-region.

The parameters that determine the grape quality, and therefore the optimal harvest time, suffer variations during berry ripening, related to climate change, with the widely known problem of the gap between technological and phenolic maturities. However, there are few studies about its incidence on grape nitrogen composition. For this reason, the use of an elicitor, methyl jasmonate (MeJ), alone or with urea, is proposed as a tool to reduce climatic decoupling, allowing to establish the harvest time in order to achieve the optimum grape quality. The aim was to study the effect of MeJ and MeJ+Urea foliar applications on the evolution of Tempranillo amino acids content throughout the grape maturation. Three treatments were foliarly applied, at veraison and 7 days later: control (water), MeJ (10 mM) and MeJ+Urea (10 mM+6 kg N/ha). Grape samples were taken at five stages of maturation: day before the first and second applications, 15 days after the second application (pre-harvest), harvest day, and 15 days after harvest (post-harvest). The amino acids analysis of the samples was carried out by HPLC. Results showed that the evolution of amino acids was similar regardless of the treatment; however, foliar applications influenced the nitrogen compounds content, i.e., there was no qualitative effect but quantitative one. Most of the amino acids reached their maximum concentration in pre-harvest, being higher in grapes from the treatments than in the control. In general, no differences in grape amino acids content were observed between MeJ and MeJ+Urea treatments. Foliar applications with MeJ and MeJ+Urea enhanced the grape amino acids content, without affecting their profile, helping to optimize their quality and allowing to establish a more complete grape ripening standard. Therefore, MeJ and MeJ+Urea foliar applications can be a simple agronomic practice, which has shown promising results in order to enhance the grape quality.

Since the 1980s global regime shift, grape growers have been steadily adapting to a changing climate. These adaptations have preserved the region-climate-cultivar rapports that have established the global trade of wine with lucrative economic benefits since the middle of 17th century. The advent of using fractions of crop and actual evapotranspiration replacement in vineyards with the use of supplemental irrigation has furthered the adaptation of wine grape cultivation. The shift in trellis systems, as well as pruning methods from positioned shoot systems to sprawling canopies, as well as adapting the bearing surface from head-trained, cane-pruned to cordon-trained, spur-pruned systems have also aided in the adaptation of grapevine to warmer temperatures. In warm climates, the use of shade cloth or over-head shade films not only have aided in arresting the damage of heat waves, but also identified opportunities to reduce the evapotranspiration from vineyards, reducing environmental footprint of vineyard. Our increase in knowledge on how best to understand the response of grapevine to climate change was aided with the identification of solar radiation exposure biomarker that is now used for phenotyping cultivars in their adaptability to harsh environments. Using fruit-based metrics such as sugar-flavonoid relationships were shown to be better indicators of losses in berry integrity associated with a warming climate, rather than solely focusing on region-climate-cultivar rapports. The resilience of wine grape was further enhanced by exploitation of rootstock × scion combinations that can resist untoward droughts and warm temperatures by making more resilient grapevine combinations. Our understanding of soil-plant-atmosphere continuum in the vineyard has increased within the last 50 years in such a manner that growers are able to use no-till systems with the aid of arbuscular mycorrhiza fungi inoculation with permanent cover cropping making the vineyard more resilient to droughts and heat waves. In premium wine grape regions viticulture has successfully adapted to a rapidly changing climate thus far, but berry based metrics are raising a concern that we may be approaching a tipping point.