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

“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.

The development of new generations of multispectral satellites such as Sentinel-2 opens possibilities as to vine water status assessment (Cohen et al., 2019). Based on a three years field campaign, a model of Stem Water Potential (SWP) estimation on vine using four satellite bands in Red, Red-Edge, NIR and SWIR domains was developed (Laroche-Pinel et al., 2021). The model relies on SWP field measures done using a pressure chamber (Scholander et al., 1965), which is a common, robust and precise method to assess vine water status (Acevedo-Opazo et al., 2008). The model was mainly developed from from SWP measures on Syrah N (Laroche Pinel E., 2021).

A large scale monitoring was organized in different vineyards in the Mediterranean region in 2021. 10 varieties amongst the most represented in this area were monitored (Cabernet sauvignon N, Chardonnay B, Cinsault N, Grenache N, Merlot N, Mourvèdre N, Sauvignon B, Syrah N, Vermentino B, Viognier B). The model was used to produce water status maps from Sentinel-2 images, starting from the beginning of June (fruit set) up to September (harvest). The average estimated SWP for each vine was compared to actual field SWP measures done by wine growers or technicians during usual monitoring of irrigation programs. The correlations between mean estimated SWP and mean measured SWP were at the same level than expected by the model. (Laroche Pinel, 2021) The general SWP kinetics were comparable. The estimated SWP would have led to same irrigation decisions concerning the date of first irrigation in comparison with measured SWP.

Acevedo-Opazo, C., Tisseyre, B., Ojeda, H., Ortega-Farias, S., Guillaume, S. (2008). Is it possible to assess the spatial variability of vine water status? OENO One, 42(4), 203.
Cohen, Y., Gogumalla, P., Bahat, I., Netzer, Y., Ben-Gal, A., Lenski, I., … Helman, D. (2019). Can time series of multispectral satellite images be used to estimate stem water potential in vineyards? In Precision agriculture ’19, The Netherlands: Wageningen Academic Publishers, pp. 445–451.
Laroche-Pinel, E., Duthoit, S., Albughdadi, M., Costard, A. D., Rousseau, J., Chéret, V., & Clenet, H. (2021). Towards vine water status monitoring on a large scale using sentinel-2 images. remote sensing, 13(9), 1837.
Laroche-Pinel,E. (2021). Suivi du statut hydrique de la vigne par télédétection hyper et multispectrale. Thèse INP Toulouse, France.
Scholander, P.F., Bradstreet, E.D., Hemmingsen, E.A., & Hammel, H.T. (1965). Sap pressure in vascular plants: Negative hydrostatic pressure can be measured in plants. Science, 148(3668), 339–346.

With contemporary climate change, cultivated Vitis vinifera L. is at risk as climate is a critical component in defining ecologically fitted plant materiel. While winegrowers can draw on the rich diversity among grapevine varieties to limit expected impacts (Morales-Castilla et al., 2020), replacing a signature variety that has created a sense of local distinctiveness may lead to several challenges. In order to sustain wine identity in uncertain climate outcomes, the study of intra-varietal diversity is important to reflect the adaptive and evolutionary potential of current cultivated varieties. The aim of this ongoing study is to understand to what extent can intra-varietal diversity be a climate change adaptation solution. With a focus on early (Sauvignon blanc, Riesling, Grolleau, Pinot noir) to moderate late (Chenin, Petit Verdot, Cabernet franc) ripening varieties, data was collected for flowering and veraison for the various studied accessions (from conservatory plots) and clones. For these phenological growing stages, heat requirements were established using nearby weather stations (adapted from the GFV model, Parker et al., 2013) and model performances were verified. Climate change projections were then integrated to predict the future behaviour of the intra-varietal diversity. Study findings highlight the strong phenotypic diversity of studied varieties and the importance of diversification to enhance climate change resilience. While model performances may require improvements, this study is the first step towards quantifying heat requirements of different clones and how they can provide adaptation solutions for winegrowers to sustain local wine identity in a global changing climate. As genetic diversity is an ongoing process through point mutations and epigenetic adaptations, perspective work is to explore clonal data from a wide variety of geographic locations.

Winemaking areas recognized as protected designations of origin (PDOs) shape important economic, environmental and cultural values that are tied to closely defined geographic locations. To preserve wine products and wine-growing practices adopted in different PDOs these areas are strictly regulated by legal specifications. However, quality viticulture is increasingly under pressure from climate change, which is altering the local conditions of many winegrowing areas. Therefore, maintaining traditional wine products will require the adoption of tailored adaptation strategies, including possible changes in the legal regulation of protected wines. To this end, it is necessary to have a comprehensive knowledge on PDOs including their extension, products and allowed practices. While there have been efforts to build databases that summarize the characteristics for individual wine PDO areas and to quantify the related effects of climate change, much information is still included only in the official documentation of the EU geographical indication register and has never been collected in a comprehensive manner. With this study we aim at filling this gap by building a spatial inventory of European wine PDOs that supports decision making in viticulture in the context of climate change. To map and characterize European wine PDOs, we analysed their legal documents and extracted relevant information useful for climate change adaptation. The output consists of a comprehensive geographical dataset that identifies the boundaries of all 1200 European wine PDOs at unprecedented spatial resolution and includes a set of legally binding regulations, such as authorized vine varieties, maximum yields and planting density. The inventory will allow researchers to analyse the impacts of climate change on European wine PDOs and support decision makers in developing tailored adaptation strategies. This includes, among others, the evaluation of new vineyard site selection, the expansion of cultivated varieties or the authorization of irrigation in vineyards.

Climate change and rising atmospheric carbon dioxide concentration is a concern for agriculture, including viticulture. Studies on elevated carbon dioxide have already been on grapevines, mainly taking place in greenhouses using potted plants or using field grown vines under higher CO2 enrichment, i.e. >650 ppm. The VineyardFACE, located at Hochschule Geisenheim University, is an open field Free Air CO2 Enrichment (FACE) experimental set-up designed to study the effects of elevated carbon dioxide using field grown vines (Vitis vinifera L. cvs. Riesling and Cabernet Sauvignon). As the carbon dioxide fumigation started in 2014, the long term effects of elevated carbon dioxide treatment can be investigated on berry ripening parameters and fruit metabolic composition.
The present study aims to investigate the effect on fruit composition under a moderate increase (+20%; eCO2) of carbon dioxide concentration, as predicted for 2050 on both Riesling and Cabernet Sauvignon. Berry composition was determined for primary (sugars, organic acids, amino acids) and secondary metabolites (anthocyanins). Special focus was given on monitoring of berry diameter and ripening rates throughout three growing seasons. Compared to previous results of the early adaptative phase of the vines [1], our results show little effect of eCO2 treatment on primary metabolites composition in berries. However, total anthocyanins concentration in berry skin was lower for eCO2 treatment in 2020, although the ratio between anthocyanins derivatives did not differ.
[1] Wohlfahrt Y., Tittmann S., Schmidt D., Rauhut D., Honermeier B., Stoll M. (2020) The effect of elevated CO2 on berry development and bunch structure of Vitis vinifera L. cvs. Riesling and Cabernet Sauvignon. Applied Science Basel 10: 2486