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…

Climate change impacts: a multi-stress issue

With the aim of producing premium wines, it is admitted that moderate environmental stresses may contribute to the accumulation of compounds of interest in grapes. However the ongoing climate change, with the appearance of more limiting conditions of production is a major concern for the wine industry economic. Will it be possible to maintain the vineyards in place, to preserve the current grape varieties and how should we anticipate the adaptation measures to ensure the sustainability of vineyards? In this context, the question of the responses and adaptation of grapevine to abiotic stresses becomes a major scientific issue to tackle. An abiotic stress can be defined as the effect of a specific factor of the physico-chemical environment of the plants (temperature, availability of water and minerals, light, etc.) which reduces growth, and for a crop such as the vine, the yield, the composition of the fruits and the sustainability of the plants. Water stress is in many minds, but a systemic vision is essential for at least two reasons. The first reason is that in natural environments, a single factor is rarely limiting, and plants have to deal with a combination of constraints, as for example heat and drought, both in time and at a given time. The second reason is that plants, including grapevine, have central mechanisms of stress responses, as redox regulatory pathways, that play an important role in adaptation and survival. Here we will review the most recent studies dealing with this issue to provide a better understanding of the grapevine responses to a combination of environmental constraints and of the underlying regulatory pathways, which may be very helpful to design more adapted solutions to cope with climate change.

Rapid damage assessment and grapevine recovery after fire

There is increasing scientific consensus that climate changeis the underlying cause of the prolonged dry and hot conditions that have increased the risk of extreme fire weather in many countries around the world. In December 2019, a bushfire event occurred in the Adelaide Hills, South Australia where 25,000 hectares were burnt and in vineyards and surrounding areas various degrees of scorching and infrastructure damage occurred. The ability to coordinate and plan recovery after a fire event relies on robust and timely data. The current practice for measuring the scale and distribution of fire damage is to walk or drive the vineyard and score individual vines based on visual observation. The process is time consuming, subjective, or semi-quantitative at best. After the December 2019 fires, it took many months to access properties and estimate the area of vineyard damaged. This study compares the rapid assessment and mapping of fire damage using high-resolution satellite imagery with more traditional ground based measures. Satellite imagery tracking vineyard recovery in the season following the bushfire is being correlated to field assessments of vineyard productivity such as canopy health and development, fertility and carbohydrate storage. Canopy health in the seasons following the fires correlated to the severity of the initial fire damage. Severely damaged vines had reduced canopy growth, were infertile or had very low fertility as well as lower carbohydrate levels in buds and canes during dormancy, which reduced productivity in the seasons following the bushfire event. In contrast, vines that received minor damage were able to recover within 1-2 years. Tools that rapidly and affordably capture the extent and severity of damage over large vineyard area will allow producers, government and industry bodies to manage decisions in relation to fire recovery planning, coordination and delivery, improving the efficiency and effectiveness of their response.

Phenolic composition of Tempranillo Blanco grapes changes after foliar application of urea

Our research aimed to determine the effect and efficiency of foliar application of urea on the phenolic composition of Tempranillo Blanco grapes. The field experiment was carried out in 2019 and 2020 seasons and the plot was located in D.O.Ca Rioja (North of Spain). The vineyard was Vitis vinifera L. Tempranillo Blanco and grafted on Richter-110 rootstock. The treatments were control (C), whose plants were sprayed with water and three doses of urea: plants were sprayed with urea 3 kg N/ha (U3), 6 kg N/ha (U6) and 9 kg N/ha (U9). The applications were performed in two phenological stages, pre-veraison (Pre) and veraison (Ver). Also, each of the treatments was repeated one week later. Control and treatments were performed in triplicate and arranged in a randomised block design. Grapes were harvested at optimum ripening stage. High-performance liquid chromatography was used to analyse the phenolic composition of the grapes. Finally, the results obtained from the analytical determinations – flavonols, flavanols and non-flavonoid (hydroxybenzoic acids, hydroxycinnamic acids and stilbenes) – were studied statistically by analysis of variance. The results showed that, in 2019, U6-Pre and U9-Pre treatments increased the hydroxybenzoic acid content in grapes, and also all foliar treatments applied at Pre enhanced the stilbene concentration. Moreover, U3-Ver was the only treatment that rose flavonol and stilbene contents in the Tempranillo Blanco grapes. In 2020, all treatments applied at Pre enhanced the flavonol concentration in grapes. Furthermore, U3-Pre and U9-Pre treatments increased stilbene content in grapes. Nevertheless, the hydroxybenzoic acid content was improved by U6-Ver and U9-Ver and besides, hydroxycinnamic acid concentration in grapes was increased by all treatments applied at Ver. In conclusion, the lower and highest dose of urea (U3 and U9), applied at pre-veraison, were the best treatments to improve the Tempranillo Blanco grape phenolic composition.

Teasing apart terroir: the influence of management style on native yeast communities within Oregon wineries and vineyards

Newer sequencing technologies have allowed for the addition of microbes to the story of terroir. The same environmental factors that influence the phenotypic expression of a crop also shape the composition of the microbial communities found on that crop. For fermented goods, such as wine, that microbial community ultimately influences the organoleptic properties of the final product that is delivered to customers. Recent studies have begun to study the biogeography of wine-associated microbes within different growing regions, finding that communities are distinct across landscapes. Despite this new knowledge, there are still many questions about what factors drive these differences. Our goal was to quantify differences in yeast communities due to management style between seven pairs of conventional and biodynamic vineyards (14 in total) throughout Oregon, USA. We wanted to answer the following questions: 1) are yeast communities distinct between biodynamic vineyards and conventional vineyards? 2) are these differences consistent across a large geographic region? 3) can differences in yeast communities be tied to differences in metabolite profiles of the bottled wine? To collect our data we took soil, bark, leaf, and grape samples from within each vineyard from five different vines of pinot noir. We also collected must and a 10º brix sample from each winery. Using these samples, we performed 18S amplicon sequencing to identify the yeast present. We then used metabolomics to characterize the organoleptic compounds present in the bottled wine from the blocks the year that we sampled. We are actively in the process of analysing our data from this study.

Diagnosis of soil quality and evaluation of the impact of viticultural practices on soil biodiversity in a vineyard in southwestern France

Viticulture is facing two major changes – climate change and agroecological transition. In both cases, soil quality is seen as a lever to move towards a more sustainable viticulture. However, soil biological quality is little considered in the implementation of viticultural practices. Gascogn’Innov (2017-2022) is an Operational Group funded by the European Innovation Partnership for Agriculture. As such, it brings together winegrowers from the south-west of France, scientists, advisors and technicians, around a project focused on viticultural soil biological functioning and the design of technical routes more respectful toward soil heritage. To achieve this, the project aims to acquire references on the impact of viticultural practices on soil biology from a dynamic way, and to test a methodology to integrate information provided by the soil bioindicators to manage farming systems. A set of indicators of soil biological quality are evaluated in the project: microorganisms (bacteria and fungi abundance and diversity), fauna (abundance and diversity of nematodes and earthworms), physico-chemical characteristics, soil structure assessment and degradation rate of organic matter. Based on a network of 13 plots that have been subject to an initial diagnosis in 2017, several agronomical practices to restore soil fertility are experimented to redesign the cropping system (for instance plant cover, organic matter inputs, reduction of herbicides, mineral fertilizers). System redesign was made in collaboration by winegrowers and an interdisciplinary group of experts (agronomists, biologists). Several indicators are measured on vine and soil at each vintage to assess vine health and productivity. At the end of the project (2021), a final diagnosis was carried out. Gascogn’Innov allowed to create a regional database on the quality of wine-growing soils, which permitted to evaluate the effect of practices according to soil types. Especially, decreasing the intensity of tillage and increasing the duration and diversity of grass coverage tends to increase the abundance of all the organisms studied. This project confirmed the value of soil biological quality indicators to drive the sustainability of practices, but also highlighted the key-role of expertise, in both agronomy and soil biology, to help winegrowers understand and appropriate their soil quality diagnoses.