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…

Assessment of the impact of actions in the vineyard and its surrounding environment on biodiversity in Rioja Alavesa (Spain)

Traditional viticulture areas have experienced in the last decades an intensification of field practices, linked to an increased use of fertilisers and phytosanitary products, and to a more intensive mechanization and uniformization of the landscape. This change in management has sometimes led to higher rates of soil erosion andloss of soil structure, fertility decline, groundwater contamination, and to an increased pressure of pests and diseases. Additionally, intensification usually leads to a simplification of landscapes, of particular concern in prestigious wine grape regions where the economical revenue encourages the conversion of land use from natural habitats to high value wine grape production. To revert this trend, it is necessary that growers implement actions that promote biodiversity in their vineyards. The aim of this study is to assess the impact of the implementation of cover crops, vegetational corridors, dry stone walls and vineyard biodiversity hotspots estimated through the study of arthropods. The work has been carried out in four vineyards in Rioja Alavesa belonging to Ostatu winery, where these infrastructures were implemented in 2020. The presence and diversity of arthropods was studied by capturing them at different times in the season and at different distances from the infrastructure using pit-fall traps in the soil and yellow, white and blue chromatic traps at the canopy level. This is a preliminary study in which all adult insects were sorted to the taxonomic level of order and Coleoptera were classified to morphospecies. The results obtained show that there is a relationship between the basic characteristics of the vineyard and the arthropods captured, with a positive effect, although also dependent on the vineyard, of the presence of infrastructure.

Comparison of imputation methods in long and varied phenological series. Application to the Conegliano dataset, including observations from 1964 over 400 grape varieties

A large varietal collection including over 1700 varieties was maintained in Conegliano, ITA, since the 1950s. Phenological data on a subset of 400 grape varieties including wine grapes, table grapes, and raisins were acquired at bud break, flowering, veraison, and ripening since 1964. Despite the efforts in maintaining and acquiring data over such an extensive collection, the data set has varying degrees of missing cases depending on the variety and the year. This is ubiquitous in phenology datasets with significant size and length. In this work, we evaluated four state-of-the-art methods to estimate missing values in this phenological series: k-Nearest Neighbour (kNN), Multivariate Imputation by Chained Equations (mice), MissForest, and Bidirectional Recurrent Imputation for Time Series (BRITS). For each phenological stage, we evaluated the performance of the methods in two ways. 1) On the full dataset, we randomly hold-out 10% of the true values for use as a test set and repeated the process 1000 times (Monte Carlo cross-validation). 2) On a reduced and almost complete subset of varieties, we varied the percentage of missing values from 10% to 70% by random deletion. In all cases, we evaluated the performance on the original values using normalized root mean squared error. For the full dataset we also obtained performance statistics by variety and by year. MissForest provided average errors of 17% (3 days) at budbreak, 14% (4 days) at flowering, 14.5% (7 days) at veraison, and 17% (3 days) at maturity. We completed the imputations of the Conegliano dataset, one of the world’s most extensive and varied phenological time series and a steppingstone for future climate change studies in grapes. The dataset is now ready for further analysis, and a rigorous evaluation of imputation errors is included.

Terroir analysis and its complexity

Terroir is not only a geographical site, but it is a more complex concept able to express the “collective knowledge of the interactions” between the environment and the vines mediated through human action and “providing distinctive characteristics” to the final product (OIV 2010). It is often treated and accepted as a “black box”, in which the relationships between wine and its origin have not been clearly explained. Nevertheless, it is well known that terroir expression is strongly dependent on the physical environment, and in particular on the interaction between soil-plant and atmosphere system, which influences the grapevine responses, grapes composition and wine quality. The Terroir studying and mapping are based on viticultural zoning procedures, obtained with different levels of know-how, at different spatial and temporal scales, empiricism and complexity in the description of involved bio-physical processes, and integrating or not the multidisciplinary nature of the terroir. The scientific understanding of the mechanisms ruling both the vineyard variability and the quality of grapes is one of the most important scientific focuses of terroir research. In fact, this know-how is crucial for supporting the analysis of climate change impacts on terroir resilience, identifying new promised lands for viticulture, and driving vineyard management toward a target oenological goal. In this contribution, an overview of the last findings in terroir studies and approaches will be shown with special attention to the terroir resilience analysis to climate change, facing the use and abuse of terroir concept and new technology able to support it and identifying the terroir zones.

Analysis of Cabernet Sauvignon and Aglianico winegrape (V. vinifera L.) responses to different pedo-climatic environments in southern Italy

Water deficit is one of the most important effects of climate change able to affect agricultural sectors. In general, it determines a reduction in biomass production, and for some plants, as in the case of grapevine, it can endorse fruit quality. The monitoring and management of plant water stress in the vineyard

Effects of organic mulches on the soil environment and yield of grapevine

Farming management practices aiming at conserving soil moisture have been developed in arid and semiarid-areas facing water scarcity problems. Organic mulching is an effective method to manipulate the crop-growing microclimate increasing crop yield by controlling soil temperature, and retaining soil moisture by reducing soil evaporation. In this sense, the effectiveness of different organic mulching materials (straw mulch and grapevine pruning debris) applied within the row of a vineyard was evaluated on the soil and on the vine in a Tempranillo vineyard located in La Rioja (Spain). Organic mulches were compared with a traditional bare soil management technique (based on the use of herbicides to avoid weed incidence). Mulching coverages favourably influenced the soil water retention throughout all the grapevine vegetative cycle. However, the soil-moisture variation was not the same under different mulching materials, being the straw mulch (SM) the one that retained more water in comparison with grapevine pruning debris (GPD) based-cover. The changes of soil moisture in the upper surface layer (0–10 cm) were highly dynamic, probably due to water vapour fluxes across the soil-atmospheric interface. However, both, SM and GPD reduced these fluctuations as compared with bare soils. A similar trend occurred with soil temperature. Both organic mulches altered soil temperature in comparison with bare soil by reducing soil temperature in summer and raising it in winter. Moreover, the same buffering effect for the temperature on the covered soil also remains in the deeper layers. To conclude, we could see that organic mulching had a positive impact on soil-moisture storage and soil temperature and the extent of this effect depends on the type of mulching materials. These changes led to higher rates of photosynthesis and stomatal conductivity compared to bare soils, also favouring crop growth and grape yields.