Macrowine 2021
IVES 9 IVES Conference Series 9 Capture depletion of grapevine DNA: an approach to advance the study of microbial community in wine

Capture depletion of grapevine DNA: an approach to advance the study of microbial community in wine

Abstract

The use of next-generation sequencing (NGS) has helped understand microbial genetics in oenology. Current studies mainly focus on barcoded amplicon NGS but not shotgun sequencing, which is useful for functional analyses. Since the high percentage of grapevine DNA conceals the microbial DNA in must, the majority of sequencing data is wasted in bioinformatic analyses. Here we present capture depletion of grapevine whole genome DNA. By using biotinylated RNA baits made from genomic grapevine libraries, the grapevine DNA is hybridized to the baits and captured by magnetic beads while the targeted microbial DNA is left and subsequently sequenced. A pilot study was carried out to mimic degraded complex materials. Using known complex microbial samples and spiking in large portion of grapevine DNA, the hybridization and depletion took place with quantification by qPCR. Different parameters, such as hybridization hours, temperatures and bait amounts were also tested for potential effects. The aim is to provide an insight into depletion of undesired grapevine DNA and to enhance the data yield in NGS studies of microbial DNA with a more cost effective way suitable for future oenological studies.

Publication date: May 17, 2024

Issue: Macrowine 2016

Type: Poster

Authors

Sarah S.T. Mak*, Anders Hansen, Christian Carøe, Jacob Enk, Jean-Marie Rouillard, M Thomas P Gilbert

*Centre for GeoGenetics

Contact the author

Tags

IVES Conference Series | Macrowine | Macrowine 2016

Citation

Related articles…

Metabolomics of grape polyphenols as a consequence of post-harvest drying: on-plant dehydration vs warehouse withering

A method of suspect screening analysis to study grape metabolomics, was developed [1]. By performing ultra-high performance liquid chromatography (UHPLC) – high-resolution mass spectrometry (HRMS) analysis of the grape extract, averaging 320-450 putative grape compounds are identified which include mainly polyphenols. Identification of metabolites is performed by a new HRMS-database of putative grape and wine compounds expressly constructed (GrapeMetabolomics) which currently includes around 1,100 entries.

Full automation of oenological fermentations and its application to the processing of must containing high sugar or acetic acid concentrations

Climate change and harvest date decisions have led to the evolution of must quality over the last decades. Increases in must sugar concentrations are among the most obvious consequences, quantitatively. Saccharomyces cerevisiae is a robust and acid tolerant organism. These properties, its sugar to ethanol conversion rate and ethanol tolerance make it the ideal production organism for wine fermentations. Unfortunately, high sugar concentrations may affect S. cerevisiae and lead to growth inhibition or yeast lysis, and cause sluggish or stuck fermentations. Even sublethal conditions cause a hyperosmotic stress response in S. cerevisiae which leads to increased formation of fermentation by-products, including acetic acid, which may exceed legal limits in some wines.

Ageing of sweet wines: oxygen evolution according to bung and barrel type

Barrel ageing is a crucial step in the wine process because it allows many changes to the wine as enrichment, colour stabilization, clarification and also a slow oxygenation. Effects of the oak barrel have to be known to prevent oxidation of the wine. The type of bung used during ageing is also a parameter to consider. Ageing sweet wines in barrel is a real challenge. These wines may need some oxygen at the beginning of ageing but they should be protected at the end of their maturation, to avoid oxidation.

Spontaneous fermentation dynamics of indigenous yeast populations and their effect on the sensory properties of Riesling

Varietal Riesling aroma relies strongly on the formation and liberation of bound aroma compounds. Floral monoterpenes, green C6-alcohols, fruity C13-norisoprenoids and spicy volatile phenols are predominantly bound to disaccharides, which are produced and stored in the grape berry during berry maturation. Grape processing aims to extract maximum amount of the precursors from the berry skin to increase the potential for a strong varietal aroma in the wine. Subsequent yeast selection plays an important part in this process.

Comprehensive two-dimensional gas chromatography coupled with Tof-MS, a powerful tool for analysis of the volatomes of grapes and wines

Comprehensive two-dimensional gas chromatography (GCxGC) has emerged as a powerful analytical technique for unraveling the volatile composition of complex matrices. This work will present three applications of GCxGC Tof-MS to the oenological field, aimed to identify novel biomarkers to be used in the quality control process of the wine industry. Comprehensive mapping of volatile compounds was conducted in a large sample of 70 sparkling wines, produced by 48 different wineries across 6 vintages and representative of the two main production areas for premium Italian sparkling wines (Franciacorta (FC) and Trentodoc (TN)), using HS-SPME followed by GCxGC-Tof-MS and multivariate analysis. Selection and identification of 196 putative biomarkers allowed clear separation of sparkling wines from FC and TN.