Macrowine 2021
IVES 9 IVES Conference Series 9 Genetic traceability of ‘Nebbiolo’ musts and wines by single nucleotide polymorphism (SNP) genotyping assays

Genetic traceability of ‘Nebbiolo’ musts and wines by single nucleotide polymorphism (SNP) genotyping assays

Abstract

AIM: ‘Nebbiolo’ (Vitis vinifera L.) is one of the most ancient and prestigious Italian grape cultivars. It is renowned for its use in producing monovarietal high-quality red wines, such Barolo and Barbaresco. Wine quality and value can be heavily modified if cultivars other than those allowed are employed. The fight against fraud to safeguard high-quality productions requires an effective varietal identification system applicable in musts and wines.

METHODS: Single-nucleotide polymorphisms (SNPs) are considered the newest type of molecular marker for grapevine identification. We developed and investigated the efficiency of SNP TaqMan® assays in the varietal authentication of ‘Nebbiolo’ musts and wines. ‘Nebbiolo’-specific SNPs were identified starting from available databases and 260 genotypes analysed by Vitis18kSNP array.

RESULTS: Only two markers (SNP_15082 and SNP_14783) were sufficient to distinguish ‘Nebbiolo’ from more than 1,100 genotypes. In experimental vinifications, these SNP TaqMan® assays correctly identified ‘Nebbiolo’ in all wine-making steps, including wines 1 year after bottling. The high sensitivity of the assays allowed identifying, for the first time, mixtures of 1% in musts at the end of maceration, blends of 10% in musts at the end of malolactic fermentation and wines contamination of 10–20% with non-‘Nebbiolo’ genotypes. In commercial wines, the amplification efficiency of these SNPs was partially limited by the low amount of grapevine DNA and the presence of PCR inhibitors in DNA extracts. However, at least one SNP amplified correctly in all the commercial wines tested.

CONCLUSIONS:

The TaqMan® genotyping assay is a rapid, highly sensitive and specific methodology with remarkable potential for varietal identification in wines.

DOI:

Publication date: September 7, 2021

Issue: Macrowine 2021

Type: Article

Authors

Giorgio Gambino, Paolo BOCCACCI, Walter CHITARRA, Anna SCHNEIDER

Institute for Sustainable Plant Protection, National Research Council (IPSP-CNR). Strada delle Cacce 73, 10135 Torino, Italy. Institute for Sustainable Plant Protection, National Research Council (IPSP-CNR). Strada delle Cacce 73, 10135 Torino, Italy. Council for Agricultural Research and Economics, Viticultural and Enology Research Centre (CREA-VE). Via XXVIII Aprile 26, 31015 Conegliano (TV), Italy. Institute for Sustainable Plant Protection, National Research Council (IPSP-CNR). Strada delle Cacce 73, 10135 Torino, Italy. Luca ROLLE, Department of Agricultural, Forest and Food Sciences, University of Torino. Largo Braccini 2, 10095 Grugliasco (TO), Italy.

Contact the author

Keywords

grapevine, musts, wines, genetic traceability, snp, blends

Citation

Related articles…

Impact on leaf morphology of Vitis vinifera L. cvs Riesling and Cabernet Sauvignon under Free Air Carbon dioxide Enrichment (FACE)

Atmospheric carbon dioxide (CO2) concentration has continuously increased since pre-industrial times from 280 ppm in 1750, and is predicted to exceed 700 ppm by the end of 21st century. For most of C3 plant species elevated CO2 (eCO2) improve photosynthetic apparatus results in an increased plant biomass production. To investigate the effects of eCO2 on morphological leaf characteristics the two Vitis vinifera L. cultivars, Riesling and Cabernet Sauvignon, grown in the Geisenheim VineyardFACE (Free Air Carbon dioxide Enrichment) system were used. The FACE site is located at Geisenheim University (49° 59′ N, 7° 57′ E, 94 m above sea level), Germany and was implemented in 2014 comparing future atmospheric CO2-concentrations (eCO2, predicted for the mid-21st century) with current ambient CO2-conditions (aCO2). Experiments were conducted under rain-fed conditions for two consecutive years (2015 and 2016). Six leaves per repetition of the CO2 treatment were sampled in the field and immediately fixed in a FAA solution (ethanol, H2O, formaldehyde and glacial acetic acid). After 24 h leaf samples were transferred and stored in an ethanol solution. Subsequently, leaf tissue was dehydrated using ethanol series and embedded in paraffin. By using a rotary microtomesections of 5 µm were prepared and fixed on microscopic slides. Subsequent the samples were stained using consecutive staining and washing solutions. Afterwards pictures of the leaf cross-sections were taken using a light microscope and consecutive measurements were conducted with an open source image software. Differences found in leaf cross-sections of the two CO2 treatments were detected for the palisade parenchyma. Leaf thickness, upper and lower epidermis and spongy parenchyma remained less affected under eCO2 conditions. The observed results within grapevine leaf tissues can provide first insights to seasonal adaptation strategies of grapevines under future elevated CO2 concentrations.

Elucidating vineyard site contributions to key sensory molecules: Identification of correlations between elemental composition and volatile aroma profile of site-specific Pinot noir wines

The reproducibility of elemental profile in wines produced across multiple vintages has been previously reported using grapes from a single scion clone of Vitis vinifera L. cv. Pinot noir. The grapevines were grown on fourteen different vineyard sites, from Oregon to southern California in the U.S.A., which span distances from approximately hundreds of meters to 1450 km, while elevations range from near sea level to nearly 500 m. In addition, sensorial (i.e. aroma, taste, and mouthfeel) and chemical (i.e. polyphenolic and volatile) differences across the different vineyard sites have also been observed among these wines at two aging time points. While strong evidence exists to support that grapes grown in different regions can produce wines with unique chemical and sensorial profiles, even when a single clone is used, the understanding of growing site characteristics that result in this reproducible differentiation continues to emerge. One hypothesis is that the elemental profile that a vineyard site imparts to the grape berries and the resulting wine is an important contributor to this differentiation in chemistry and sensory of wines. For example, various classes of enzymes that catalyze the formation of key aroma compounds or their precursors require specific metals. In this work, we begin to report correlations between elemental and volatile aroma profiles of site-specific Pinot noir wines, made under standardized winemaking conditions, that have been previously shown to be distinguished separately by these chemical analyses.

The combined effects of climate, soils, and deficit irrigation on yield and quality of Touriga Nacional under high atmospheric demand in the Douro Region

Global warming is one of the biggest environmental, social and economic threats in several viticultural regions. In the Douro Valley, changes are expected in the coming years, namely an increase in temperature and a decrease in precipitation. These changes are likely to have consequences for the production and quality of wine.
The aim of this study was to explore the effects of different soil characteristics combined with several deficit irrigation strategies, managed throughout ETc references and predawn leaf water potentials thresholds, on physiology, yield, and qualitative attributes on the Touriga Nacional variety under years of mild to severe water and heat stress.
The studies were conducted over seven years (2015 to 2021) in two plots of a commercial vineyard located at Quinta do Ataíde (Symington Family Estates) planted in 2011 and 2014 at 170 meters elevation, growing under three water regimes: non-irrigated (NI) and two deficit irrigation strategies (30% and 60% ETc) assessed weekly by Ψpd. The site has an annual rainfall below 500 mm, with high atmospheric demand. Climate data was collected from a weather station, located on site. Berry ripening was followed weekly for fruit analysis. At harvest, yield, vigour and pruning weight per vine were determined from 90 vines by treatment. Each season at veraison the NDVI Index was accessed by a drone. The soils physic-chemistry in the experimental blocs were analysed and grouped by SWHC. Delta C-13 analyses were also performed per treatment in two years.Irrigation had a positive effect on yield per vine, mostly due to an increase in berry and cluster weight, and fertility index through the years. A significant increase in sugar content, colour and phenols was observed with deficit irrigation in some years, but vine vigour related to soil characteristics had by far the greatest impact on quality.

Simulating climate change impact on viticultural systems in historical and emergent vineyards

Global climate change affects regional climates and hold implications for wine growing regions worldwide. Although winegrowers are constantly adapting to internal and external factors, it seems relevant to develop tools, which will allow them to better define actual and future agro-climatic potentials. Within this context, we develop a modelling approach, able to simulate the impact of environmental conditions and constraints on vine behaviour and to highlight potential adaptation strategies according to different climate change scenarios. Our modeling approach, named SEVE (Simulating Environmental impacts on Viticultural Ecosystems), provides a generic modeling framework for simulating grapevine growth and berry ripening under different conditions and constraints (slope, aspect, soil type, climate variability…) as well as production strategies and adaptation rules according to climate change scenarios. Each activity is represented by an autonomous agent able to react and adapt its reaction to the variability of environmental constraints. Using this model, we have recently analyzed the evolution of vineyards’ exposure to climatic risks (frost, pathogen risk, heat wave) and the adaptation strategies potentially implemented by the winegrowers. This approach, implemented for two climate change scenarios, has been initiated in France on traditional (Loire Valley) and emerging (Brittany) vineyards. The objective is to identify the time horizons of adaptations and new opportunities in these two regions. Carried out in collaboration with wine growers, this approach aims to better understand the variability of climate change impacts at local scale in the medium and long term.

A blueprint for managing vine physiological balance at different spatial and temporal scales in Champagne

In Champagne, the vine adaptation to different climatic and technical changes during these last 20 years can be seen through physiological balance disruptions. These disruptions emphasize the general grapevine decline. Since the 2000s, among other nitrogen stress indicators, the must nitrogen has been decreasing. The combination of restricted mineral fertilizers and herbicide use, the growing variability of spring rainfall, the increasing thermal stress as well as the soil type heterogeneity are only a few underlying factors that trigger loss of physiological balance in the vineyards. It is important to weigh and quantify the impact of these factors on the vine. In order to do so, the Comité Champagne uses two key-tools: networking and modelization. The use of quantitative and harmonized ecophysiological indicators is necessary, especially in large spatial scales such as the Champagne appellation. A working group with different professional structures of Champagne has been launched by the Comité Champagne in order to create a common ecophysiology protocol and thus monitor the vine physiology, yearly, around 100 plots, with various cultural practices and types of soil. The use of crop modelling to follow the vine physiological balance within different pedoclimatic conditions enables to understand the present balance but also predict the possible disruptions to come in future climatic scenarios. The physiological references created each year through the working group, benefit the calibration of the STICS model used in Champagne. In return, the model delivers ecophysiology indicators, on a daily scale and can be used on very different types of soils. This study will present the bottom-up method used to give accurate information on the impacts of soil, climate and cultural practices on vine physiology.